EP0626387A1 - Nucleosides and oligonucleotides with 2'-ether groups - Google Patents

Abstract

Compounds of the formula <IMAGE> in which R1 and R2 are, independently of one another, hydrogen or a protective group, or R1 has these meanings and R2 is a radical forming a phosphorus-containing nucleotide bridging group; B is a purine or pyrimidine residue or an analogue thereof; and R3 is a radical of the formula Ia, Ib or Ic <IMAGE> in which R4 is hydrogen, C1-C21-alkyl, C2-C21-alkenyl, C2-C21- alkynyl or -C(=O)-alkyl; R5 is hydrogen, C1-C10-alkyl, -CH2-O-R6 or a radical of the formula Ib; R6 is hydrogen, C1-C22-alkyl, C3-C21-alkenyl, partially or completely fluorine-substituted C1-C10-alkyl or -[(CH2)2-O]m-R7; R 7 is hydrogen or C1-C21-alkyl; Z is -(CH2)p- or -(CH2-CH-O)q-CH2 CH2-, where Z can in the case of -CH2- be unsubstituted or substituted by one or more C1-C10-alkyl, C5-C6-cycloalkyl or unsubstituted or C1-C4-alkyl-substituted phenyl, independently of one another; n is a number from 1 to 12; m is a number from 1 to 4; p is a number from 1 to 10; and q is a number from 1 to 4, are described.

Description

The invention relates to ribo-nucleoside analogs whose 2'-OH group is etherified with polyol derivatives, a process for their preparation, oligonucleotides with these nucleosides and the use of the nucleosides for the production of oligonucleotides with the same or different nucleoside units in the molecule.

Nucleosides and oligonucleotides have found broad interest as antiviral agents or because of their ability to interact with nucleic acids ("antisense" oligonucleotides) and the biological activity associated with them, see for example Uhlmann, E., Peyman, A., Chemical Reviews 90: 543-584 (1990). To provide nucleosides with new properties or to improve the interaction of antisense oligonucleotides with natural nucleic acids and their stability towards nucleases, the sugar residues of nucleosides (or the nucleotide units in oligonucleotides) or the internucleotide phosphate bond in oligonucleotides have been modified in a wide variety of ways, see, for example, Marquez, VE, Lim, MI, Medicinal Research Reviews 6: 1-40 (1986), Hélène, C., Toulmé, JJ, Biochimica et Biophysica Acta 1049: 99-125 (1990), English, U., Gauss, DH, Angewandte Chemie 103: 629-646 (1991), Matteucci, MD, Bischofberger, N., Annual Reports in Medicinal Chemistry 26: 87-296 (1991). Cook, PD, Anti-Cancer Drug Design 6: 585-607 (1991) and WO 91/06556 describe nucleosides which are modified on the 2'-OH group of the sugar. The modifications described lead to increased nuclease resistance; the longer the alkyl radical, the higher the nuclease resistance. A small increase in binding affinity is observed with small alkyl residues such as methyl, ethyl or propyl, but with longer chains the binding affinity drops drastically. Nucleosides with polyol derivatives as side chains on the 2'-OH group are unknown and have never been incorporated into oligonucleotides. Surprisingly, the modifications according to the invention increase the binding affinity for the complementary RNA not only with shorter chains but also with longer chains. Based on the published data, this result was not to be expected. In analogy to the 2'-OH-modified oligoribonucleotides, the compounds according to the invention are also distinguished by a striking nuclease resistance. In addition, oligonucleotides which the nucleosides according to the invention contain an increased cellular uptake and consequently have an improved bioavailability and activity in vivo.

worin R₁ und R₂ unabhängig voneinander für Wasserstoff oder eine Schutzgruppe stehen oder R₁ diese Bedeutungen hat und R₂ für einen eine phosphorhaltige Nukleotid-Brückengruppe bildenden Rest steht; B einen Purin- oder Pyrimidinrest oder ein Analoges davon bedeutet; und R₃ einen Rest der Formel Ia, Ib oder Ic bedeutet

worin

R₄

Wasserstoff, C₁-C₂₁-Alkyl, C₂-C₂₁-Alkenyl, C₂-C₂₁-Alkinyl oder -C(=O)-Alkyl;

R₅

Wasserstoff, C₁-C₁₀-Alkyl, -CH₂-O-R₆ oder einen Rest der Formel Ib;

R₆

Wasserstoff, C₁-C₂₂-Alkyl, C₃-C₂₁-Alkenyl, teilweise oder vollständig mit Fluor substituiertes C₁-C₁₀-Alkyl oder -[(CH₂)₂-O]_m-R₇;

R₇

Wasserstoff oder C₁-C₂₁-Alkyl;

Z

-(CH₂)_p- oder -(CH₂-CH₂-O)_q-CH₂CH₂-, wobei Z im Fall von -CH₂- unsubstituiert oder mit einem oder mehreren C₁-C₁₀-Alkyl, C₅-C₆-Cycloalkyl oder unsubstituiertem oder mit C₁-C₄-Alkyl substituiertem Phenyl unabhängig voneinander substituiert sein kann;

n

eine Zahl von 1 bis 12;

m

eine Zahl von 1 bis 4;

p

eine Zahl von 1 bis 10; und

q

eine Zahl von 1 bis 4 bedeuten;

wobei R₄ für den Fall n=1 und R₅=Wasserstoff nicht Wasserstoff bedeutet.The invention relates to compounds of the formula I.

wherein R₁ and R₂ independently represent hydrogen or a protecting group or R₁ has these meanings and R₂ stands for a residue forming a phosphorus-containing nucleotide bridge group; B represents a purine or pyrimidine residue or an analog thereof; and R₃ represents a radical of formula Ia, Ib or Ic

wherein

R₄

Hydrogen, C₁-C₂₁ alkyl, C₂-C₂₁ alkenyl, C₂-C₂₁ alkynyl or -C (= O) alkyl;

R₅

Hydrogen, C₁-C₁₀ alkyl, -CH₂-O-R₆ or a radical of the formula Ib;

R₆

Hydrogen, C₁-C₂₂-alkyl, C₃-C₂₁-alkenyl, partially or completely fluorine-substituted C₁-C₁₀-alkyl or - [(CH₂) ₂-O] _m -R₇;

R₇

Hydrogen or C₁-C₂₁ alkyl;

Z.

- (CH₂) _p - or - (CH₂-CH₂-O) _q -CH₂CH₂-, where Z in the case of -CH₂- is unsubstituted or with one or more C₁-C₁₀ alkyl, C₅-C₆ cycloalkyl or unsubstituted or with C₁ -C₄-alkyl substituted phenyl may be independently substituted;

n

a number from 1 to 12;

m

a number from 1 to 4;

p

a number from 1 to 10; and

q

represent a number from 1 to 4;

where R₄ for the case n = 1 and R₅ = hydrogen is not hydrogen.

In a preferred embodiment, R₄ is hydrogen, C₁-C₂₁-alkyl, C₂-C₂₁-alkenyl or C₂-C₂₁-alkynyl. In a more preferred embodiment, R₄ is H, C₁-C₈-alkyl, C₂-C₈-alkenyl or C₂-C₈-alkynyl. In a particularly preferred embodiment, R₄ is H or C₁-C₄alkyl. Examples of R₄ are methyl, ethyl, n- or i-propyl, n, i- and t-butyl, the isomers of pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl , Heptadecyl, octadecyl, nonadecyl, undecyl, vinyl, allyl, but-3-en-1-yl, but-2-en-1-yl, pentenyl, octenyl, dodecenyl, octadecenyl, ethinyl, prop-1-in-1 -yl, prop-1-in-3-yl, but-1-in-1- or-4-yl. Particularly preferred R₄ radicals are H, methyl, ethyl, n- or i-propyl or n-butyl. The numerical values n are preferably 1 to 8, particularly preferably 1 to 6, in particular 1 to 3.

In a preferred embodiment, R₅ is hydrogen, C₁-C₅-alkyl or -CH₂-O-R₆, particularly preferably hydrogen, methyl or -CH₂-OH, -CH₂-O-C₁-C₂₂-alkyl or -CH₂-O - [( CH₂) ₂-O] _m -C₁-C₁₀-alkyl, where m is a number from 1 to 4.

In a preferred embodiment, R₁ and R₂ represent hydrogen.

Protective groups and methods for derivatizing the hydroxyl groups with such protective groups are generally known in sugar and nucleotide chemistry and are described, for example, by Greene, BT, Protective Groups in Organic Synthesis, Wiley Interscience, New York (1991), by Sonveaux, E., Bioorganic Chemistry 14: 274-325 (1986) or by Beaucage, SL, Iyer, R., Tetrahedron 48: 2223-2311 (1992). Examples of such protective groups are: benzyl, methylbenzyl, dimethylbenzyl, methoxybenzyl, dimethoxybenzyl, bromobenzyl, 2,4-dichlorobenzyl; Diphenylmethyl, Di (methylphenyl) methyl, Di (dimethylphenyl) methyl, Di (methoxyphenyl) methyl, Di (dimethoxyphenyl) methyl, Triphenylmethyl, Tris-4,4 ', 4''- tert.butylphenylmethyl, Di-p-anisylphenylmethyl, Tri (methylphenyl) methyl, tri (dimethylphenyl) methyl, methoxyphenyl (diphenyl) methyl, di (methoxyphenyl) phenylmethyl, tri (methoxyphenyl) methyl, tri (dimethoxyphenyl) methyl; Triphenylsilyl, alkyldiphenylsilyl, dialkylphenylsilyl and trialkylsilyl with 1 to 20, preferably 1 to 12 and particularly preferably 1 to 8 C atoms in the alkyl groups, for example trimethylsilyl, triethylsilyl, tri-n-propylsilyl, i-propyldimethylsilyl, t-butyl -dimethylsilyl, t-butyl-diphenylsilyl, n-octyl-dimethylsilyl, (1,1,2,2-tetramethylethyl) -dimethylsilyl; - (C₁-C₈-alkyl) ₂Si-O-Si (C₁-C₈-alkyl) ₂-, wherein alkyl means, for example, methyl, ethyl n- and i-propyl, n-, i- or t-butyl; C₂-C₁₂-, especially C₂-C₈ acyl, such as acetyl, propanoyl, butanoyl, pentanoyl, hexanoyl, benzoyl, methylbenzoyl, methoxybenzoyl, chlorobenzoyl and bromobenzoyl; R _S1 -SO₂-, wherein R _{S1 is} C₁-C₁₂-alkyl, especially C₁-C₆-alkyl, C₅- or C₆-cycloalkyl, phenyl, benzyl, C₁-C₁₂- and especially C₁-C₄-alkylphenyl, or C₁-C₁₂- and especially C₁-C₄ alkylbenzyl, or halophenyl or halobenzyl, for example methyl, ethyl, propyl, butyl, phenyl, benzyl, p-bromo, p-methoxy and p-methylphenylsulfonyl; unsubstituted or substituted by F, Cl, Br, C₁-C₄-alkoxy, tri- (C₁-C₄-alkyl) silyl or C₁-C₄-alkylsulfonyl-substituted C₁-C₁₂-, preferably C₁-C₈-alkoxycarbonyl, for example methoxy, ethoxy -, n- or i-Propoxy- or n-, i- or t-butoxycarbonyl, 2-trimethylsilylethoxycarbonyl, 2-methylsulfonylethoxycarbonyl, allyloxycarbonyl or unsubstituted or substituted for alkoxycarbonyl or phenyloxycarbonyl or benzyloxycarbonyl, for example methyl- or methoxy- or chlorophenyloxycarbonyl -benzyloxycarbonyl, and 9-fluorenylmethyloxycarbonyl. If R₁ and / or R₂ are alkyl, it can be substituted with F, Cl, Br, C₁-C₄alkoxy, phenyloxy, chlorophenyloxy, methoxyphenyloxy, benzyloxy, methoxybenzyloxy or chlorophenyloxy. R₁ and R₂ in formula I can be the same or different protecting groups.

In a particularly preferred embodiment, R₁ and R₂ represent benzyl, methylbenzyl, dimethylbenzyl, methoxybenzyl, dimethoxybenzyl, halogenated benzyl, especially bromobenzyl; Diphenylmethyl, di (methylphenyl) methyl, di (dimethylphenyl) methyl, di (methoxyphenyl) methyl, di (methoxyphenyl) (phenyl) methyl, triphenylmethyl, tris-4,4 ', 4' '- tert.butylphenylmethyl, di-p anisylphenylmethyl, tri (methylphenyl) methyl, tri (dimethylphenyl) methyl, tri (methoxyphenyl) methyl, tri (dimethoxyphenyl) methyl; Trimethylsilyl, triethylsilyl, tri-n-propylsilyl, i-propyldimethylsilyl, t-butyl-dimethylsilyl, t-butyl-diphenylsilyl, n-octyl-dimethylsilyl, (1,1, 2,2-tetramethylethyl) -dimethylsilyl, - (CH₃) ₂Si-O-Si (CH₃) ₂-, - (i-C₃H₇) ₂Si-O-Si (i-C₃H₇) ₂-; Acetyl, propanoyl, butanoyl, pentanoyl, hexanoyl, benzoyl, methylbenzoyl, methoxybenzoyl, chlorobenzoyl and bromobenzoyl; Methyl, ethyl, propyl, butyl, phenyl, benzyl, p-bromo, p-methoxy and p-methylphenylsulfonyl; Methoxy-, ethoxy-, n- or i-propoxy- or n-, i- or t-butoxycarbonyl, or phenyloxycarbonyl, benzyloxycarbonyl, methyl- or methoxy- or chlorophenyloxycarbonyl or -benzyloxycarbonyl or 9-fluorenylmethyloxycarbonyl. R₁ and R₂ mean advantageously the same protective groups.

entsprechen, worin Y_a Wasserstoff, C₁-C₁₂-Alkyl, C₆-C₁₂-Aryl, C₇-C₂₀-Aralkyl, C₇-C₂₀-Alkaryl, -OR_b, -SR_b, -NH₂, Primäramino, Sekundäramino, O^⊖M^⊕ oder S^⊖M^⊕ darstellt; X_a Sauerstoff oder Schwefel bedeutet; R_a Wasserstoff, M^⊕, C₁-C₁₂-Alkyl, C₂-C₁₂-Alkenyl, C₆-C₁₂-Aryl, oder die Gruppe R_aO- für N-Heteroaryl-N-yl mit 5 Ringgliedern und 1 bis 3 N-Atomen steht; R_b Wasserstoff, C₁-C₁₂-Alkyl oder C₆-C₁₂-Aryl bedeutet; und M^⊕ für Na^⊕, K^⊕, Li^⊕, NH₄^⊕ steht oder Primär-, Sekundär-, Tertiär- oder Quartenärammonium darstellt; wobei Alkyl, Aryl, Aralkyl und Alkaryl in Y_a, R_a und R_b unsubstituiert oder mit Alkoxy, Alkylthio, Halogen, -CN, -NO₂, Phenyl, Nitrophenyl, oder Halogenphenyl substituiert ist.R₂ can as a phosphorus containing a nucleotide bridge group forming radical of the formula P1 or P2

correspond in which Y _{a is} hydrogen, C₁-C₁₂-alkyl, C₆-C₁₂-aryl, C₇-C₂₀-aralkyl, C₇-C₂₀-alkaryl, -OR _b , -SR _b , -NH₂, primary amino, secondary ^amino , O ^⊖ M ^⊕ or S ^{⊖ represents} M ^⊕ ; X _{a represents} oxygen or sulfur; R _{a is} hydrogen, M ^⊕ , C₁-C₁₂-alkyl, C₂-C₁₂-alkenyl, C₆-C₁₂-aryl, or the group R _a O- for N-heteroaryl-N-yl with 5 ring members and 1 to 3 N atoms stands; R _{b represents} hydrogen, C₁-C₁₂ alkyl or C₆-C₁₂ aryl; and M ^{⊕ represents} Na ^⊕ , K ^⊕ , Li ^⊕ , NH₄ ^⊕ or represents primary, secondary, tertiary or quaternary ammonium; wherein alkyl, aryl, aralkyl and alkaryl in Y _a , R _a and R _{b are} unsubstituted or substituted with alkoxy, alkylthio, halogen, -CN, -NO₂, phenyl, nitrophenyl, or halophenyl.

Y _a preferably contains 1 to 12 and particularly preferably 1 to 6 carbon atoms as primary amino, and preferably 2 to 12 and particularly preferably 2 to 6 carbon atoms as secondary amino.

The primary amino and secondary amino can be, for example, radicals of the formula R _c R _d N, in which R _{c is} H or independently has the meaning of R _d , and R _{d is} C₁-C₂₀-, preferably C₁-C₁₂- and particularly preferably C₁-C₆-alkyl, C₁-C₂₀-, preferably C₁-C₁₂- and particularly preferably C₁-C₆-aminoalkyl, C₁-C₂₀-, preferably C₁-C₁₂- and particularly preferably C₁-C₆-hydroxyalkyl; Carboxyalkyl or carbalkoxyalkyl, the carbalkoxy group containing 2 to 8 carbon atoms and the alkyl group 1 to 6, preferably 1 to 4 carbon atoms; C₂-C₂₀-, preferably C₂-C₁₂- and particularly preferably C₂-C₆-alkenyl; Phenyl, mono- or di- (C₁-C₄ alkyl or alkoxy) phenyl, benzyl, mono- or di- (C₁-C₄ alkyl or alkoxy) benzyl; or 1,2-, 1,3- or 1,4-imidazolyl-C₁-C₆-alkyl, or R _c and R _d together tetra- or pentamethylene, 3-oxa-1,5-pentylene, -CH₂-NR _{e represent} -CH₂CH₂- or -CH₂CH₂-NR₁₉-CH₂CH₂-, wherein R _{e represents} H or C₁-C₄-alkyl. The amino group in the aminoalkyl can be substituted with one or two C₁-C₄ alkyl or hydroxyalkyl groups. The hydroxyl group in the hydroxyalkyl can be etherified with C₁-C₄-alkyl.

Primary, secondary, tertiary and quaternary ammonium means for Y _a in connection with the definition of M ^⊕ an ion of the formula R _f R _g R _h R _i N ^⊕ , in which R _f C₁-C₂₀-, preferably C₁-C₁₂- and particularly preferably C₁-C₆-alkyl, -aminoalkyl, -hydroxyalkyl; Carboxyalkyl or carbalkoxyalkyl, the carbalkoxy group containing 2 to 8 carbon atoms and the alkyl group 1 to 6, preferably 1 to 4 carbon atoms; C₂-C₂₀-, preferably C₂-C₁₂- and particularly preferably C₂-C₆-alkenyl; Phenyl, mono- or di- (C₁-C₄ alkyl or alkoxy) phenyl, benzyl, mono- or di- (C₁-C₄ alkyl or alkoxy) benzyl; or 1,2-, 1,3- or 1,4-imidazolyl-C₁-C₆-alkyl, and R _g , R _h and R _{i are} independently hydrogen or have the meaning of R _f , or R _f and R _g together represent tetra- or pentamethylene, 3-oxa-1,5-pentylene, -CH₂-NR _e -CH₂CH₂- or -CH₂CH₂-NR₂-CH₂CH₂-, wherein R _{e represents} H or C₁-C₄-alkyl, and R _h and R _i independently of one another have the meaning of R _f . The amino group in the aminoalkyl can be substituted with one or two C₁-C₄ alkyl or hydroxyalkyl groups. The hydroxyl group in the hydroxyalkyl can be etherified with C₁-C₄-alkyl.

Examples of carboxyalkyl are carboxymethyl, carboxyethyl, carboxypropyl and carboxybutyl, and examples of carbalkoxyalkyl are these carboxyalkyl groups esterified with methyl or ethyl. Examples of alkenyl are allyl, but-1-en-3-yl or -4-yl, pent-3- or 4-en-1-yl or -2-yl, hex-3- or -4- or -5 -en-1-yl or -2-yl. Examples of alkyl- and alkoxyphenyl or -benzyl are methylphenyl, dimethylphenyl, ethylphenyl, diethylphenyl, methylbenzyl, dimethylbenzyl, ethylbenzyl, diethylbenzyl, methoxyphenyl, dimethoxyphenyl, ethoxyphenyl, diethoxyphenyl, methoxybenzyl, dimethoxybenzyl, ethoxybenzyl, ethoxybenzyl, ethoxybenzyl. Examples of imidazolylalkyl, in which the alkyl group preferably contains 2 to 4 carbon atoms, are 1,2-, 1,3- or 1,4-imidazolylethyl or -n-propyl or -n-butyl. R₁₉ preferably represents H, methyl or ethyl.

Preferred examples of primary amino and secondary amino are methyl, ethyl, dimethyl, diethyl, di-i-propyl, mono- or di- (1-hydroxy-eth-2-yl) -, phenyl- and benzylamino, acetylamino and Benzoylamino and piperidinyl, piperazinyl and morpholinyl.

Preferred examples of primary and secondary ammonium are methyl, ethyl, dimethyl, diethyl, di-i-propyl, mono- or di- (1-hydroxy-eth-2-yl), phenyl and benzylammonium.

Examples of Y _a , R _a and R _b as alkyl are methyl, ethyl and the isomers of propyl, butyl, pentyl, hexyl, heptyl and octyl; Examples of Y _a , R _a and R _b as aryl are phenyl and naphthyl; Examples of R _a as alkenyl are allyl and (C₁-C₄-alkyl) CH = CH-CH₂-; Examples of Y _a as aralkyl are phenyl-C _n H _2n - with n equal to a number from 1 to 6, particularly Benzyl; Examples of Y _a as alkaryl are mono-, di- and tri (C₁-C₄-alkyl) phenyl. Preferred substituents are chlorine, bromine, methoxy, -NO₂, -CN, 2,4-dichlorophenyl and 4-nitrophenyl. Examples of R _b are 2,2,2-trichloroethyl, 4-chlorophenyl, 2-chlorophenyl and 2,4-dichlorophenyl; and examples of R _b O- as N-heteroaryl are pyrrol-N-yl, triazol-N-yl and benzotriazol-N-yl.

In a particularly preferred embodiment, R _{a is} β-cyanoethyl and Y _{a is} di (i-propylamino).

worin R_b1 für H, Cl, Br, OH oder -O-C₁-C₁₂-Alkyl steht, und R_b2, R_b3 und R_b5 unabhängig voneinander H, OH, SH, NH₂, NHNH₂, NHOH, NHO-C₁-C₁₂-Alkyl, -N=CH-N(C₁-C₁₂-Alkyl)₂, -N=CH-N-Cycloalkyl, F, Cl, Br, C₁-C₁₂-Alkyl,- Hydroxy-C₁-C₁₂-alkyl, Amino-C₁-C₁₂-alkyl, C₁-C₁₂-Alkoxy, Benzyloxy, C₁-C₁₂-Alkylthio, wobei die Hydroxyl- und Aminogruppen unsubstituiert oder mit einer Schutzgruppe substituiert sind, Phenyl, Benzyl, Primäramino mit 1 bis 20 C-Atomen oder Sekundäramino mit 2 bis 30 C-Atomen bedeuten, R_b4 Wasserstoff, CN oder -C≡C-R_b7, R_b6 und R_b7 Wasserstoff oder C₁-C₄-Alkyl darstellen.If B represents a purine residue or an analogue thereof, it can be a residue of the formula II, IIa, IIb, IIc, IId, IIe or IIf,

wherein R _{b1 is} H, Cl, Br, OH or -O-C₁-C₁₂-alkyl, and R _b2 , R _b3 and R _b5 independently of one another H, OH, SH, NH₂, NHNH₂, NHOH, NHO-C₁-C₁₂ -Alkyl, -N = CH-N (C₁-C₁₂-alkyl) ₂, -N = CH-N-cycloalkyl, F, Cl, Br, C₁-C₁₂-alkyl, - hydroxy-C₁-C₁₂-alkyl, amino- C₁-C₁₂-alkyl, C₁-C₁₂-alkoxy, benzyloxy, C₁-C₁₂-alkylthio, where the hydroxyl and amino groups are unsubstituted or substituted with a protective group, phenyl, benzyl, primary amino with 1 to 20 C atoms or secondary amino with 2 to 30 carbon atoms, R _{b4 are} hydrogen, CN or -C≡CR _b7 , R _b6 and R _{b7 are} hydrogen or C₁-C₄-alkyl.

Suitable protecting groups have been mentioned previously. Preferred protecting groups are C₁-C₈ acyl groups, such as acetyl, propionyl, butyroyl and benzoyl. R _b6 preferably represents H or methyl.

The primary amino preferably contains 1 to 12 and particularly preferably 1 to 6 carbon atoms, and the secondary amino preferably 2 to 12 and particularly preferably 2 to 6 carbon atoms.

Some examples of alkyl, alkoxy, alkylthio, hydroxyalkyl and aminoalkyl, which preferably contain 1 to 6 carbon atoms, are methyl, ethyl and the isomers of propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl and dodecyl , as well as corresponding alkoxy, alkylthio, hydroxyalkyl and aminoalkyl radicals. The alkyl, alkoxy, alkylthio, hydroxyalkyl and aminoalkyl particularly preferably contains 1 to 4 carbon atoms. Preferred alkyl, alkoxy, alkylthio, hydroxyalkyl and aminoalkyl radicals are methyl, ethyl, n- and i-propyl, n-, i- and t-butyl, methoxy, ethoxy, methylthio and ethylthio, aminomethyl, aminoethyl, Hydroxymethyl and hydroxyethyl.

The primary amino and secondary amino can be, for example, radicals of the formula R _a1 R _a2 N, in which R _{a1 is} H or independently has the meaning of R _a2 , and R _{a2 is} C₁-C₂₀-, preferably C₁-C₁₂- and particularly preferably C₁-C₆ alkyl, aminoalkyl, hydroxyalkyl; Carboxyalkyl or carbalkoxyalkyl, the carbalkoxy group containing 2 to 8 carbon atoms and the alkyl group 1 to 6, preferably 1 to 4 carbon atoms; C₂-C₂₀-, preferably C₂-C₁₂- and particularly preferably C₂-C₆-alkenyl; Phenyl, mono- or di- (C₁-C₄ alkyl or alkoxy) phenyl, benzyl, mono- or di- (C₁-C₄ alkyl or alkoxy) benzyl; or 1,2-, 1,3- or 1,4-imidazolyl-C₁-C₆-alkyl, or R _a1 and R _a2 together tetra- or pentamethylene, 3-oxa-1,5-pentylene, -CH₂-NR _{represent a3} -CH₂CH₂- or -CH₂CH₂-NR _a3 -CH₂CH₂-, wherein R _{a3 represents} H or C₁-C₄-alkyl. The amino group in the aminoalkyl can have one or two C₁-C₄ alkyl or hydroxyalkyl groups be substituted. The hydroxyl group in the hydroxyalkyl can be etherified with C₁-C₄-alkyl.

Examples of alkyl have been given previously. Examples of aminoalkyl are aminomethyl, aminoethyl, 1-aminoprop-2-yl or -3-yl, 1-amino-but-2-yl or -3-yl or -4-yl, N-methyl- or N, N- Dimethyl or N-ethyl or N, N-diethyl or N-2-hydroxyethyl or N, N-di-2-hydroxyethylaminomethyl or -aminoethyl or -aminopropyl or -aminobutyl. Examples of hydroxyalkyl are hydroxymethyl, 1-hydroxy-eth-2-yl, 1-hydroxy-prop-2- or -3-yl, 1-hydroxy-but-2-yl, -3-yl or -4-yl. Examples of carboxyalkyl are carboxymethyl, carboxyethyl, carboxypropyl and carboxybutyl, and examples of carbalkoxyalkyl are these carboxyalkyl groups esterified with methyl or ethyl. Examples of alkenyl are allyl, but-1-en-3-yl or -4-yl, pent-3-or 4-en-1-yl or -2-yl, hex-3- or -4- or -5 -en-1-yl or -2-yl. Examples of alkyl and alkoxyphenyl or benzyl are methylphenyl, dimethylphenyl, ethylphenyl, diethylphenyl, methylbenzyl, dimethylbenzyl, ethylbenzyl, diethylbenzyl, methoxyphenyl, dimethoxyphenyl, ethoxyphenyl, diethoxyphenyl, methoxybenzyl, dimethoxybenzyl, ethoxybenzyl, ethoxybenzyl. Examples of imidazolylalkyl, in which the alkyl group preferably contains 2 to 4 carbon atoms, are 1,2-, 1,3- or 1,4-imidazolylethyl or -n-propyl or -n-butyl. R _a3 preferably represents H, methyl or ethyl.

Preferred examples of primary amino and secondary amino are methyl, ethyl, dimethyl, diethyl, allyl, mono- or di- (1-hydroxy-eth-2-yl) -, phenyl- and benzylamino, acetylamino, isobutyrylamino and benzoylamino .

In a preferred embodiment, R _{b1 represents} hydrogen. In another preferred embodiment, R _{b5 represents} hydrogen. In a further preferred embodiment, R _b2 and R _b3 independently of one another denote H, F, Cl, Br, OH, SH, NH₂, NHOH, NHNH₂, methylamino, dimethylamino, benzoylamino, isobutyrylamino, methoxy, ethoxy and methylthio.

In addition to purine, xanthine, hypoxanthine, adenine, N-methyladenine, N-benzoyladenine, 2-methylthioadenine, 2-aminoadenine, 6-hydroxypurine, 2-amino-6-chloropurine, 2-amino-6-methylthiopurine are some examples of analogs of the purine series , Guanine, N-isobutyrylguanine. Adenine, 2-aminoadenine and guanine and their base-protected derivatives are particularly preferred.

worin R_b6 H oder C₁-C₄-Alkyl und R_b8 H, OH, SH, NH₂, NHNH₂, NHOH, NHO-C₁-C₁₂-Alkyl, -N=CH-N(C₁-C₁₂-Alkyl)₂, -N=CH-N-Cycloalkyl, F, Cl, Br, C₁-C₁₂-Alkyl, Hydroxy-C₁-C₁₂-alkyl, Amino-C₁-C₁₂-alkyl, C₁-C₁₂-Alkoxy, Benzyloxy, C₁-C₁₂-Alkylthio, wobei die Hydroxyl- und Aminogruppen unsubstituiert oder mit einer Schutzgruppe substituiert sind, Phenyl, Benzyl, Primäramino mit 1 bis 20 C-Atomen, Sekundäramino mit 2 bis 30 C-Atomen, C₁-C₁₂-Alkenyl oder C₁-C₁₂-Alkinyl bedeuten, und die NH₂-Gruppe in Formel IIIb unsubstituiert oder mit C₁-C₆-Alkyl, Benzoyl oder einer Schutzgruppe substituiert ist, sowie die Dihydroderivate der Reste der Formeln III, IIIa, IIIb und IIIc. Bevorzugt stellt R_b8 in Formel III H, C₁-C₆-Alkyl oder -Hydroxyalkyl, C₂-C₆-Alkenyl oder -Alkinyl, F, Cl, Br, NH₂, Benzoylamino, Mono- oder Di-C₁-C₆-alkylamino dar. Bevorzugt stellt R_b8 in Formel IIIb und IIIc H, C₁-C₆-Alkyl oder -Alkoxy oder -Hydroxyalkyl, C₂-C₆-Alkenyl oder -Alkinyl, F, Cl, Br, NH₂, Benzoylamino, Mono- oder Di-C₁-C₆-alkylamino dar.If B in formula I represents a pyrimidine residue, it is preferably an uracil, thymine or cytosine residue of the formula III, IIIa, IIIb or IIIc

wherein R _b6 H or C₁-C₄-alkyl and R _b8 H, OH, SH, NH₂, NHNH₂, NHOH, NHO-C₁-C₁₂-alkyl, -N = CH-N (C₁-C₁₂-alkyl) ₂, -N = CH-N-cycloalkyl, F, Cl, Br, C₁-C₁₂-alkyl, hydroxy-C₁-C₁₂-alkyl, amino-C₁-C₁₂-alkyl, C₁-C₁₂-alkoxy, benzyloxy, C₁-C₁₂-alkylthio, where the hydroxyl and amino groups are unsubstituted or substituted by a protective group, phenyl, benzyl, primary amino having 1 to 20 carbon atoms, secondary amino having 2 to 30 carbon atoms, C₁-C₁₂ alkenyl or C₁-C₁₂ alkynyl, and the NH₂ group in formula IIIb is unsubstituted or substituted by C₁-C₆-alkyl, benzoyl or a protective group, and the dihydro derivatives of the radicals of the formulas III, IIIa, IIIb and IIIc. R _b8 in formula III is preferably H, C₁-C₆ alkyl or hydroxyalkyl, C₂-C₆ alkenyl or alkynyl, F, Cl, Br, NH₂, benzoylamino, mono- or di-C₁-C₆-alkylamino. Preferred R _b8 in formula IIIb and IIIc represents H, C₁-C₆-alkyl or alkoxy or -hydroxyalkyl, C₂-C₆-alkenyl or -alkynyl, F, Cl, Br, NH₂, benzoylamino, mono- or di-C₁-C₆- alkylamino.

R _b6 preferably represents H or methyl. R _b8 in formula III preferably denotes H, F, Cl, Br, NH₂, NHCH₃, N (CH₃) ₂, C₁-C₄-alkyl, C₂-C₄-alkenyl or C₂-C₄-alkyn-1-yl. R _b8 in formula IIIb and IIIc is preferably H, C₁-C₄-alkyl, especially methyl, C₂-C₄-alkenyl, especially vinyl or C₂-C₄-alkyn-1-yl, especially 1-propin-1-yl, or NH₂ , NHCH₃ or (CH₃) ₂N.

Some examples of pyrimidine analogs are uracil, thymine, cytosine, 5-fluorouracil, 5-chlorouracil, 5-bromouracil, dihydrouracil, 5-methylcytosine, 5-propinthymine and 5-propynecytosine.

worin R₁ und R₂ unabhängig voneinander für Wasserstoff oder eine Schutzgruppe stehen oder R₁ diese Bedeutungen hat und R₂ für einen eine phosphorhaltige Nukleotid-Brückengruppe bildenden Rest steht; B einen Purin- oder Pyrimidinrest oder ein Analoges davon bedeutet; und

• (a) R₃ einen Rest der Formel Ia bedeutet
  
  worin R₄ Wasserstoff, C₁-C₂₁-Alkyl, C₂-C₂₁-Alkenyl, C₂-C₂₁-Alkinyl oder -C(=O)-Alkyl;
  R₅ Wasserstoff, C₁-C₁₀-Alkyl, -CH₂-O-R₆ oder einen Rest der Formel Ib; R₆ Wasserstoff, C₁-C₂₂-Alkyl, C₃-C₂₁-Alkenyl, teilweise oder vollständig mit Fluor substituiertes C₁-C₁₀-Alkyl oder -[(CH₂)₂-O]_m-R₇; R₇ Wasserstoff oder C₁-C₂₁-Alkyl; n eine Zahl von 1 bis 12; und m eine Zahl von 1 bis 4 bedeuten; wobei R₄ für den Fall n=1 und R₅=Wasserstoff nicht Wasserstoff ist,
  das dadurch gekennzeichnet ist, dass man eine Verbindung der Formel IVa
  
  worin R₁₄ und R₁₅ für gleiche oder verschiedene Schutzgruppen stehen und B einen Purin- oder Pyrimidinrest oder ein Analoges davon bedeutet, wobei funktionelle Gruppen im Basenrest B durch Schutzgruppen geschützt sind, in einem inerten Lösungsmittel mit einer Verbindung der Formel A umsetzt
  
  worin R₄, R₅ und n die oben genannten Bedeutungen haben und X Cl, Br, I, Tosyl-O oder Mesyl-O bedeutet;
• (b) R₃ einen Rest der Formel Ic bedeutet
  
  das dadurch gekennzeichnet ist, dass man eine Verbindung der Formel IVa in einem inerten Lösungsmittel mit einer Verbindung der Formel B umsetzt
  
  worin X Cl, Br, I, Tosyl-O oder Mesyl-O bedeutet;
• (c) R₃ einen Rest der Formel Ib bedeutet
  
  worin R₅ Wasserstoff, C₁-C₁₀-Alkyl, -CH₂-O-R₆ oder einen Rest der Formel Ib; R₆ Wasserstoff, C₁-C₂₂-Alkyl, C₃-C₂₁-Alkenyl, teilweise oder vollständig mit Fluor substituiertes C₁-C₁₀-Alkyl oder -[(CH₂)₂-O]_m-R₇; R₇ Wasserstoff oder C₁-C₂₁-Alkyl; Z -(CH₂)_p- oder -(CH₂-CH-O)_q-CH₂CH₂-, wobei Z im Fall von -CH₂- unsubstituiert oder mit einem oder mehreren C₁-C₁₀-Alkyl, C₅-C₆-Cycloalkyl oder unsubstituiertem oder mit C₁-C₄-Alkyl substituiertem Phenyl unabhängig voneinander substituiert sein kann; m eine Zahl von 1 bis 4; p eine Zahl von 1 bis 10; und q eine Zahl von 1 bis 4 bedeuten;
  das dadurch gekennzeichnet ist, dass man das in (b) erhaltene Epoxid in Gegenwart von H₂O und BF₃ öffnet und mit einer Verbindung der Formel X'-(CH₂)_q-X' oder X'-(CH₂-CH-O)_r-X', worin X' Cl, Br, I, Tosyl-O oder Mesyl-O bedeutet, zu einem neuen Ring schliesst;
• (d) R₃ einen Rest der Formel Ia darstellt,
  das dadurch gekennzeichnet ist, dass man das in (b) erhaltene Epoxid in Gegenwart von R₆OH und BF₃ umsetzt und gegebenenfalls die freiwerdende Hydroxylgruppe in einen Ether oder Ester überführt;
• (e) R₃ einen Rest der Formel Ia darstellt,
  das dadurch gekennzeichnet ist, dass man das in (b) erhaltene Epoxid in Gegenwart von BF₃ und beispielsweise NaBH₄ hydriert und gegebenenfalls die freiwerdende Hydroxylgruppe in einen Ether oder Ester überführt;
• (f) R₃ einen Rest der Formel Ia darstellt,
  das dadurch gekennzeichnet ist, dass man das in (b) erhaltene Epoxid mit einem entsprechenden Grignard-Reagenz umsetzt und gegebenenfalls die freiwerdende Hydroxylgruppe in einen Ether oder Ester überführt; oder
• g) R₃ einen Rest der Formel Ia, Ib oder Ic bedeutet,
  das dadurch gekennzeichnet ist, dass man eine Verbindung der Formel IVb
  
  worin R₁₄ und R₁₅ die oben erwähnte Bedeutung haben und A für eine Abgangsgruppe, bevorzugt Alkoxy, Acyloxy, Mesyl-O, Tosyl-O und besonders bevorzugt OCH₃, OCOCH₃ und Benzoyloxy steht, nach einem der in (a) bis (f) beschriebenen Verfahren an der 2'-OH Gruppe substituiert und dann in an sich bekannter Weise den Basenrest B durch Substitution einführt [E. Lukevics, A. Zablocka, Nucleoside Synthesis, Ellis Horwood, New York (1991)]; gegebenenfalls die Schutzgruppen R₁₄ und R₁₅ abspaltet und den die phosphorhaltige Nukleotid-Brückengruppe bildenden Rest einführt.

The invention further relates to a process for the preparation of compounds of the formula I,

wherein R₁ and R₂ independently represent hydrogen or a protecting group or R₁ has these meanings and R₂ stands for a residue forming a phosphorus-containing nucleotide bridge group; B represents a purine or pyrimidine residue or an analog thereof; and

• (a) R₃ is a radical of the formula Ia
  
  wherein R₄ is hydrogen, C₁-C₂₁ alkyl, C₂-C₂₁ alkenyl, C₂-C₂₁ alkynyl or -C (= O) alkyl;
  R₅ is hydrogen, C₁-C₁₀ alkyl, -CH₂-O-R₆ or a radical of the formula Ib; R₆ is hydrogen, C₁-C₂₂-alkyl, C₃-C₂₁-alkenyl, partially or completely fluorine-substituted C₁-C₁₀-alkyl or - [(CH₂) ₂-O] _m -R₇; R₇ is hydrogen or C₁-C₂₁ alkyl; n is a number from 1 to 12; and m represents a number from 1 to 4; where R₄ is not hydrogen for the case n = 1 and R₅ = hydrogen,
  which is characterized in that a compound of formula IVa
  
  wherein R₁₄ and R₁₅ represent the same or different protective groups and B is a purine or pyrimidine radical or an analog thereof, functional groups in the base radical B being protected by protective groups, reacted in an inert solvent with a compound of the formula A.
  
  wherein R₄, R₅ and n have the meanings given above and X is Cl, Br, I, tosyl-O or mesyl-O;
• (b) R₃ is a radical of the formula Ic
  
  which is characterized in that a compound of the formula IVa is reacted in an inert solvent with a compound of the formula B.
  
  wherein X is Cl, Br, I, Tosyl-O or Mesyl-O;
• (c) R₃ is a radical of the formula Ib
  
  wherein R₅ is hydrogen, C₁-C₁₀ alkyl, -CH₂-O-R₆ or a radical of the formula Ib; R₆ is hydrogen, C₁-C₂₂-alkyl, C₃-C₂₁-alkenyl, partially or completely fluorine-substituted C₁-C₁₀-alkyl or - [(CH₂) ₂-O] _m -R₇; R₇ is hydrogen or C₁-C₂₁ alkyl; Z - (CH₂) _p - or - (CH₂-CH-O) _q -CH₂CH₂-, where Z in the case of -CH₂- is unsubstituted or with one or more C₁-C₁₀-alkyl, C₅-C₆-cycloalkyl or unsubstituted or with C₁-C₄ alkyl substituted phenyl may be independently substituted; m is a number from 1 to 4; p is a number from 1 to 10; and q represents a number from 1 to 4;
  which is characterized in that the epoxy obtained in (b) in the presence of H₂O and BF₃ opens and with a compound of the formula X '- (CH₂) _q -X' or X '- (CH₂-CH-O) _r -X', where X 'Cl, Br, I, tosyl-O or mesyl -O means closes to a new ring;
• (d) R₃ represents a radical of the formula Ia,
  which is characterized in that the epoxide obtained in (b) is reacted in the presence of R₆OH and BF₃ and optionally the released hydroxyl group is converted into an ether or ester;
• (e) R₃ represents a radical of the formula Ia,
  which is characterized in that the epoxide obtained in (b) is hydrogenated in the presence of BF₃ and, for example, NaBH₄ and optionally the released hydroxyl group is converted into an ether or ester;
• (f) R₃ represents a radical of the formula Ia,
  which is characterized in that the epoxide obtained in (b) is reacted with a corresponding Grignard reagent and, if appropriate, the hydroxyl group released is converted into an ether or ester; or
• g) R₃ is a radical of the formula Ia, Ib or Ic,
  which is characterized in that a compound of formula IVb
  
  wherein R₁₄ and R₁₅ have the meaning mentioned above and A is a leaving group, preferably alkoxy, acyloxy, mesyl-O, tosyl-O and particularly preferably OCH₃, OCOCH₃ and benzoyloxy, by one of the processes described in (a) to (f) substituted on the 2'-OH group and then introduces the base radical B by substitution in a manner known per se [E. Lukevics, A. Zablocka, Nucleoside Synthesis, Ellis Horwood, New York (1991)]; optionally splitting off the protective groups R₁₄ and R₁₅ and introducing the residue forming the phosphorus-containing nucleotide bridge group.

Compounds of the formula I with R₅ equal to hydrogen and n = 1 are also obtained by reacting a compound of the formula IVa with a compound of the formula X-CH₂-CO-OR if the CO group is subsequently reduced to a CH₂OH group and this then alkylated.

The compounds of the formulas IVa and IVb, A and B are known, some are commercially available or can be prepared by known or analogous processes.

Inert solvents are, for example, hydrocarbons, halogenated hydrocarbons, alkylated carboxamides and lactams, ethers, nitriles such as acetonitrile, dialkyl sulfones or sulfoxides or cyclic sulfones and sulfoxides.

The reaction temperatures in process steps (a) to (g) are between -50 to 200 ° C, preferably between 0 to 90 ° C.

The reactions are advantageously carried out in the presence of bases, for example alkali metal hydrides, alcoholates, hydroxides, carbonates, trialkylamines or diazabicycloundecene.

The reduction can be carried out, for example, catalytically or using borohydrides.

The isolation of the compounds of formula I and their purification is carried out by methods known per se, such as precipitation or crystallization and filtration, and chromatographic methods.

Oligonucleotides can be built up from the compounds of formula I, which, because of their interaction with nucleic acids, have valuable biological activities and can be used as active pharmaceutical ingredients or as diagnostics.

The invention further relates to the use of the compounds of the formula I for the preparation of oligonucleotides which contain the same or different monomer units of compounds of the formula I, but at least one monomer unit of compounds of the formula I in combination with monomer units of other natural or synthetic nucleosides, wherein the oligonucleotides contain 2 to 200 monomer units. The oligonucleotides preferably contain 2 to 100, particularly preferably 2 to 50, and particularly preferably 4 to 30, monomer units. Oligonucleotides which contain the same or different monomer units of compounds of the formula I are preferred. Oligonucleotides which additionally contain monomer units of synthetic or natural nucleosides which are derived from D-ribose or 2-deoxyribose are also preferred.

bedeutet, und B und R₃ die für die Verbindungen der Formel I angegebenen Bedeutungen haben einschliesslich der Bevorzugungen und Beispiele.The invention further relates to oligonucleotides of the formula V.

5'-U- (OYOV-) _x OYOW-3 '(V),


wherein x is a number from 0 to 200 and Y is a nucleotide bridge group, U, V and W each represent the same or different radicals of natural or synthetic nucleosides and at least one of the radicals U, V and / or W is a radical of the formula VI

means, and B and R₃ have the meanings given for the compounds of formula I including the preferences and examples.

A preferred bridging group Y is the group -P (O) O ^⊖ - occurring in natural oligonucleotides. Examples of further bridging groups are -P (O) S ^⊖ -, -P (S) S ^⊖ -, -P (O) R₁₆-, P (O) NR₁₇R₁₈, or -CH₂-, where R₁₆ is H or C₁-C₆- Represents alkyl and R₁₇ and R₁₈ independently of one another have the meaning of R₁₆. In formula V, x preferably stands for a number from 0 to 100, particularly preferably for a number from 1 to 50 and particularly preferably for a number from 3 to 29. The radicals of the formula VI can be bound at the end or in the nucleotide sequence, all of which or several, for example 2 to 5, residues of the formula VI can follow one another, or the residues of the formula VI can be bound between residues of natural or synthetic nucleosides, or there can be mixed forms of these distributions in the nucleotide sequence.

A very particularly preferred embodiment is oligonucleotides of the formula V, in which x represents a number from 2 to 50, preferably 2 to 30, Y represents the group -P (O) O ^⊖ -, U, V and W are each the same or mean different residues of a natural nucleoside and at least one of the residues U, V or W corresponds to formula VI. Suitable natural nucleosides are adenosine, cytidine, guanosine, uridine, 2-aminoadenine, 5-methylcytosine, 2'-deoxyadenosine, 2'-deoxycytidine, 2'-deoxyguanosine and thymidine. Adenine, cytosine, guanine, thymine and uracil are particularly worth mentioning as natural nucleoside bases. The residues of the formula VI can be bonded terminally or in the nucleotide sequence, it being possible for all or more, for example 2 to 5, identical or different residues of the formula VI to follow one another, or identical or different residues of the formula VI are bound between residues of natural nucleosides , or mixed forms of these distributions in the nucleotide sequence. In another preferred embodiment of oligonucleotides of the formula V, all radicals U, V and W correspond to the same or different radicals of the formula VI. X preferably represents a number from 3 to 29 and preferably contains a total of 1 to 12 radicals of the formula VI.

und die Verbindung der Formel D mit Hilfe eines "linkers", zum Beispiel Bernsteinsäureanhydrid, an ein festes Trägermaterial bindet, zum Beispiel an Controlled Pore Glass (CPG), das langkettige Alkylaminogruppen enthält. In einem separaten Verfahren wird die Hydroxylgruppe der Verbindung der Formel D derivatisiert, zum Beispiel zu einem Phosphoramidit unter Verwendung von R'OP[N(i-Propyl)₂)]₂ zu einer Verbindung der Formel E

wobei R' zum Beispiel β-Cyanoethyl darstellt.The oligonucleotides according to the invention can be prepared in a manner known per se by various methods in optionally automated DNA synthesizers which can be purchased together with procedural instructions. In the case of the bridging group -P (O) O ^⊖ -, for example, the phosphortriester process, the phosphite triester process or the H-phosphonate process can be used, which are familiar to the person skilled in the art. In the phosphite triester process, for example, one can proceed in such a way that the nucleosides of the formula I, in which R₁ and R₂ each denote H, are reacted with a protective group agent, for example 4,4'-dimethoxytriphenylmethyl chloride, to give a nucleoside of the formula D.

and binds the compound of formula D to a solid support material, for example to Controlled Pore Glass (CPG), which contains long-chain alkylamino groups, using a "linker", for example succinic anhydride. In a separate process, the hydroxyl group of the compound of formula D is derivatized, for example to a phosphoramidite using R'OP [N (i-propyl) ₂)] ₂ to a compound of formula E.

where R 'represents, for example, β-cyanoethyl.

After splitting off the protective group, such as the DMT group of the material bound to the support, coupling -N (i-C₃H₇) ₂ is coupled with the compound of the formula D, blocking any free hydroxyl groups (capping) and then oxidizing the formed phosphite to phosphate. After deprotection of the dimer, the reaction cycle is repeated with a compound of the formula E until an oligomer with the desired number of monomer units has been synthesized, and the product is detached from the support material. In this way, oligonucleotides are obtained in which all residues U, V and W according to formula V consist of residues of formula VI. In this way it is also possible to produce oligonucleotides with any monomer units in any sequence, depending on the use of synthetic, natural and nucleoside building blocks according to the invention in the individual reaction cycles.

The compounds of formula I according to the invention, in which R₁ and R₂ each denote H, have antiviral and antiproliferative properties and can accordingly be used as medicaments. The oligonucleotides according to the invention also have a high stability against degradation by nucleases. Their excellent pairing with complementary nucleic acid strands, especially of the RNA type, is particularly surprising. In addition, they show an unexpectedly high cellular uptake. The oligonucleotides according to the invention are therefore particularly suitable for antisense technology, ie for inhibiting the expression of undesired protein products by binding to suitable complementary nucleotide sequences of mRNA (EP 266,099, WO 87/07300 and WO 89/08146). They can be used to treat infections and diseases, for example by blocking the expression of bioactive proteins at the nucleic acid level (for example oncogenes). The oligonucleotides according to the invention are also suitable as diagnostics and can be used as gene probes for the detection of viral infections or genetically caused diseases be used by selective interaction at the level of single or double stranded nucleic acids ("gene probes"). In particular - due to the increased stability towards nucleases - a diagnostic application is possible not only in vitro but also in vivo (for example tissue samples, blood plasma and blood serum). Such possible uses are described, for example, in WO 91/06556.

Another object of the invention relates to the use of the oligonucleotides according to the invention as diagnostics for the detection of viral infections or genetically caused diseases.

Another object of the invention also relates to the nucleosides of the formula I and the oligonucleotides of the formula V for use in a therapeutic method for the treatment of diseases in warm-blooded animals, including humans, by inactivating nucleotide sequences in the body. The dose when administered to warm-blooded animals of approximately 70 kg body weight can be, for example, 0.01 to 1000 mg per day. Administration is preferably in the form of pharmaceutical preparations parenterally, for example intravenously or intraperitoneally.

The invention further relates to a pharmaceutical preparation containing an effective amount of a nucleoside of the formulas I or an oligonucleotide of the formulas V alone or together with other active ingredients, a pharmaceutical carrier material preferably in a significant amount and optionally auxiliaries.

The pharmacologically active nucleosides and oligonucleotides according to the invention can be used in the form of parenterally administrable preparations or infusion solutions. Such solutions are preferably isotonic aqueous solutions or suspensions, these being able to be prepared before use, for example in the case of lyophilized preparations which contain the active substance alone or together with a carrier material, for example mannitol. The pharmaceutical preparations can be sterilized and / or contain auxiliaries, for example preservatives, stabilizers, wetting agents and / or emulsifiers, solubilizers, salts for regulating the osmotic pressure and / or buffers. The pharmaceutical preparations, which, if desired, can contain further pharmacologically active substances such as, for example, antibiotics, are produced in a manner known per se, for example by means of conventional solution or lyophilization processes, and contain about 0.1% to 90%, in particular about 0 , 5% to about 30%, for example 1% to 5% active substance (s).

The following examples illustrate the invention. The 1 H-NMR spectra are based on the numbering of the carbon atoms in the following cyclic carbon skeletons:

Output connections:

Nucleosides (examples):

DMF

Dimethylformamid

ClBnCl₂

2,4-Dichlorbenzylchlorid

Bn

Benzyl

Ac

Acetyl

φ

Phenyl

BSA

N,N-Bistrimethylsilylacetamid

DBU

Diazabicyclo[5.4.0]undec-7-en

BOM-Cl

Benzyloxymethylchlorid

DMTCl

4,4'-Dimethoxytritylchlorid

THF

Tetrahydrofuran

Abbreviations used in the text and in the formulas:

DMF

Dimethylformamide

ClBnCl₂

2,4-dichlorobenzyl chloride

Bn

Benzyl

Ac

Acetyl

φ

Phenyl

BSA

N, N-bistrimethylsilylacetamide

DBU

Diazabicyclo [5.4.0] undec-7-ene

BOM-Cl

Benzyloxymethyl chloride

DMTCl

4,4'-dimethoxytrityl chloride

THF

Tetrahydrofuran

A) Preparation of nucleoside analogs Example A1:

28.0 g of 1-methylribose are added dropwise at 60 ° C. to an initial charge of 13.5 g of NaH in 130 ml of DMF. After the end of the H₂ development, 110.0 g of ClBnCl₂ are added dropwise. The reaction mixture is stirred at 25 ° C for 16 hours. In order to destroy any NaH still present, methanol is carefully added dropwise and the reaction mixture is then poured onto ice / water. The lumpy precipitate is filtered off and washed well with acetonitrile. Compound (A1) is obtained.

1 H-NMR (250 MHz, CDCl₃): The HC (1) proton appears as a singlet at 5.0 ppm.
MS: 638 (M⁺)

Example A2:

65.9 g of the product prepared in Example A1 are dissolved in 600 ml of methylene chloride and cooled to 0 ° C. Then dropwise 121 ml of SnCl₄ in 800 ml of methylene chloride and allowed to stand at 3 ° C. After 26 hours another 2 ml of SnCl₄ are added. After a total of 35 hours, the reaction solution is carefully poured onto 700 ml of a saturated NaHCO₃ solution. After dilution with 400 ml of methylene chloride, the Sn-containing precipitate is filtered off. The organic phase of the filtrate is dried with MgSO₄ and evaporated to the compound (A2).

1 H-NMR (250 MHz, CDCl₃): The H-C (1) proton appears at 4.90 ppm as a doublet with J = 5 Hz.

Example A3:

125.9 g of the product obtained in Example A2 are dissolved in 1 l of pyridine. 25.5 g acetic anhydride and 1 g 4-dimethylaminopyridine are added at 20 ° C. The mixture is then stirred for 17 hours. The reaction mixture is taken up in 1 l of water, acidified with concentrated hydrochloric acid and extracted with ethyl acetate. The extract is dried with MgSO₄ and evaporated. Finally, the residue is crystallized with hexane. The connection (A3) is obtained.

1 H-NMR (250 MHz, CDCl₃): 5.15 [d, J = 4.5 Hz, H -C (1)]; 3.50 (s, OC H ₃); 2.17 (s, OCOC H ₃);
MS: 522 (M⁺)
[α] _{Na (D)} = 87.4 ± 1.0 °, CHCl₃ (0.998%)

Example A4 :

24 g of thymine are slurried in 100 ml of 1,2-dichloroethane. After adding 116.4 g of BSA, the mixture is heated under reflux until a clear solution is obtained. The mixture is then cooled to 50 ° C. and 50 g of the product prepared in Example A3 and 27.5 g of trimethylsilyl trifluoromethanesulfonate are added. It is stirred for 20 hours at 70 ° C and then poured onto 300 ml of NaHCO₃ solution and stirred. After extraction with dichloroethane, it is dried with MgSO₄ and evaporated. Finally, the residue is crystallized with methanol. The connection (A4) is obtained.

1 H-NMR (250 MHz, CDCl₃): 8.25 (s, N H ; 6.10 [d, J = 4.5 Hz, H -C (1 ')]; 2.13 (s, OCOC H ₃ ); 1.66 (s, C H ₃)
MS: 616 (M⁺)

Example A5:

85 g of the product prepared in Example A4 are suspended in 850 ml of acetonitrile. 24.2 g DBU and 24.9 g BOM-Cl are added dropwise at room temperature. After stirring for 20 hours, the reaction mixture is poured into water and with ethyl acetate extracted. The extract is dried with MgSO₄ and evaporated. The connection (A5) is obtained.

1 H-NMR (250 MHz, CDCl₃): 6.05 [d, J = 4.5 Hz, H -C (1 ')]; 5.5 (AB, C H₂ ); 5.37 [dd, HC (2 ')]; 2.13 (s, OCOC H ₃); 1.55 (s, C H ₃)
MS: 736 (M⁺)

Example A6

106 g of the product produced in Example A5 are suspended in 1 l of THF. 26 g of a 30% NaOCH₃ / CH₃OH solution are added dropwise. After stirring for 2.5 hours, the reaction solution is poured into water, saturated aqueous sodium chloride solution is added and the mixture is extracted with ethyl acetate. After drying with MgSO₄, the extract is evaporated. Compound (A6) is obtained.

1 H-NMR (250 MHz, CDCl₃): 5.93 [d, J = 5 Hz, H -C (1 ')]; 5.5 (AB, C H₂ ); 3.03 (d, J = 6.5 Hz, O H ); 1.72 (s, C H ₃)
MS: 694 (M⁺)

Example A7

20.3 g of the product obtained in Example A6 are dissolved in 200 ml of THF. After adding 0.73 g of NaH, the mixture is boiled for 30 minutes. 2.89 g of 2-chloroethyl methyl ether are then added and the mixture is boiled for a further 24 hours. A further 0.5 g of NaH and 1.7 g of 2-chloroethyl methyl ether are then added and the mixture is boiled further. After a total of 32 hours, the reaction mixture is poured onto water and extracted with ethyl acetate. The extract is dried with MgSO₄ and evaporated. The residue is chromatographed on silica gel with toluene / ethyl acetate (4: 1). Compound (A7) is obtained.

1 H-NMR (250 MHz, CDCl₃): 7.65 [s, H -C (6)]; 5.93 [s, H -C (1 ')]; 5.5 (s, C H₂ ); 3.33 (s, OC H ₃); 1.6 (s, C H ₃).
MS: 752 (M⁺).

Example A8

79.4 g of the product obtained in Example A6 are dissolved in 800 ml of THF. After adding 3.3 g of NaH, the mixture is boiled briefly and then 21 g of methyl bromoacetate are added dropwise at 40 ° C. The reaction mixture is stirred at 60 ° C. for a total of 27 hours, 1 g of NaH and 2 ml of methyl bromoacetate being added after 16 hours and after 20 hours. Finally the reaction mixture is poured onto water and extracted with ethyl acetate. The extract is dried with MgSO₄ and evaporated. This gives compound (A8).

1 H-NMR (250 MHz, CDCl₃): 7.70 [s, H -C (6)]; 5.92 [s, H -C (1 ')]; 5.48 (AB, C H₂ ); 3.75 (s, OC H ₃); 1.58 (s, C H ₃).
MS: 766 (M⁺).

Example A9

• a) 37 g of the product obtained according to Example A8 are dissolved in 400 ml of THF. 1.5 g of LiBH₄ are added in portions at 20 ° C. and the mixture is stirred for 1 hour. The reaction mixture is then carefully poured onto 500 ml of water and neutralized with 32 ml of 2N aqueous hydrochloric acid. After extraction with ethyl acetate and evaporation, compound (A9) is obtained.
  
  
  1 H-NMR (250 MHz, CDCl₃): 7.65 [s, H -C (6)]; 5.96 [s, H -C (1 ')]; 5.50 (AB, C H₂ ); 2.57 (broad s, O H ); 1.60 (s, C H ₃).
  MS: 738 (M⁺).
• b) The compound (A9) is methylated with CH₃J in the presence of NaH in THF at 70 ° C. After 8 hours the mixture is worked up as described in Example A7. Compound (A7) is obtained.

Example A10

20.0 g of the product prepared in Example A7 are dissolved in 200 ml of THF and hydrogenated over 2 g of Pd / C (5%) at 25 ° C and under normal pressure for 4.5 hours (H₂ uptake 102%). After filtration and evaporation of the filtrate, the residue is dissolved in 170 ml of methanol and adjusted to a pH of 11 with a 30% NaOCH₃ / CH₃OH solution. After 24 hours, the mixture is poured onto 250 ml of water, acidified with 2N aqueous hydrochloric acid and extracted with ethyl acetate. The extract is dried with MgSO₄ and evaporated. Compound (A10) is obtained.

1 H-NMR (250 MHz, CDCl₃): 8.75 (s, N H ); 7.65 [s, H -C (6)]; 5.97 [s, H -C (1 ')]; 3.33 (s, OC H ₃); 1.60 (s, C H ₃).
MS: 632 (M⁺).

Example A11

15 g of the product prepared in Example A10 are hydrogenated in 250 ml of methanol in the presence of 9.25 g of anhydrous sodium acetate over 5 g of Pd / C (5%) at 50 ° C. and under normal pressure. After 46 hours, the hydrogenation mixture is filtered and evaporated. To remove salts, the residue is chromatographed on a small frit using silica gel (ethyl acetate / methanol 9: 1). Compound (A11) is obtained.

1 H-NMR (250 MHz, CDCl₃): 11.5 (s, N H ); 7.95 [s, H -C (6)]; 6.00 [d, J = 6 Hz, H -C (1 ')]; 5.33 (broad s, O H ); 5.20 (d, O H ); 3.32 (s, OC H ₃); 1.92 (s, C H ₃).
MS: 316 (M⁺).

Example A12

0.45 g of the product prepared in Example A11 is taken up twice in pyridine and evaporated. It is taken up again in 18 ml of pyridine and the following is added in succession: 0.2 g of triethylamine, 0.55 g of DMTCl and 25 mg of 4-dimethylamino-pyridine. After 16 hours of stirring at room temperature, the reaction mixture is diluted with 50 ml of ethyl acetate and poured onto 50 ml of water. The organic Phase is dried with MgSO₄ and concentrated. The residue is chromatographed on silica gel (hexane / ethyl acetate / triethylamine 86: 10: 4). Compound (A12) is obtained.

1 H-NMR (250 MHz, CDCl₃): 7.62 [s, H -C (6)]; 6.02 [d, J = 4 Hz, H -C (1 ')]; 3:38 (s, OC H ₃); 1.36 (s, C H ₃).

Example A13

330 mg of the product obtained in Example A12 are added to a template of 110 mg of diisopropylammonium tetrazolide, 178 mg of 2-cyanoethyl-N, N, N ', N'-tetraisopropylphosphorodiamidite and 6 ml of methylene chloride. The reaction mixture is stirred for 17 hours at room temperature and then poured onto a saturated aqueous NaHCO₃ solution. The organic phase is dried with MgSO₄ and evaporated. The residue is chromatographed on silica gel (ethanol / ethyl acetate 4: 1 with 1% addition of triethylamine). The foam obtained is dissolved in 1 ml of methyl t-butyl ether and added dropwise at 0 ° C. in pentane. Compound (A13) (diastereoisomers, 1: 1) is obtained.

1 H-NMR (250 MHz, CDCl₃): 7.70 [s, H -C (6)] and 7.63 [s, H -C (6)]; 6.08 [d, J = 4 Hz, H -C (1)]; 6.02 [d, J = 4 Hz, H -C (1 ')].

Example 14:

24.2 g of the product obtained in Example A2 are dissolved in 250 ml of THF. 8.76 g of methyl bromoacetate and 1.38 g of NaH are added, and the reaction mixture is then stirred at 60 ° C. for 3.5 hours. After adding 0.14 g of NaH and 0.53 ml of methyl bromoacetate again, the mixture is stirred at 60 ° C. for a further 3 hours. The suspension is then poured onto 300 ml of water, neutralized with 2N aqueous hydrochloric acid and extracted with ethyl acetate. The extract is dried over MgSO₄ and evaporated. Compound (A14) is obtained.

1 H-NMR (250 MHz, CDCl₃): 5.05 [d, J = 4 Hz, H -C (1 ')]; 3.50 [s, OC H ₃] and 3.75 [s, OC H ₃].
MS: 552 (M⁺).

Example A15

5.0 g of the product obtained in Example A14 are dissolved in 60 ml of acetonitrile. Now 4.88 g of bisilylated thymine are added and 2.6 g of trimethylsilyl trifluoromethanesulfonate are added dropwise at 60 ° C. with stirring. After stirring for 3.5 hours, the reaction solution is cooled, poured onto a saturated aqueous NaHCO₃ solution and stirred. Now it is extracted with ethyl acetate, the organic phase is dried with MgSO₄ and evaporated. The residue is chromatographed on silica gel (toluene / ethyl acetate 1: 1). Compound (A15) is obtained.

1 H-NMR (250 MHz, CDCl₃): 8.36 (s, N H ); 7.70 [s, H -C (6)]; 5.94 [d, J = 1.5 Hz, H -C (1 ')]; 3.73 (s, OC H ₃); 1.58 [s, C H ₃].
MS: 646 (M⁺).
b) The reaction with benzyloxymethyl chloride gives compound (A8).

Example A16

Analogous to the above working instructions, further 2'-OH modifications are carried out and listed in Table 1.

Example A17

5.0 g of the product prepared in Example A6 are dissolved in 50 ml of THF. After adding 0.21 g of NaH, the reaction mixture is heated to 60 ° C. for 45 minutes, then mixed with 1.57 g of diethylene glycol chloroethyl methyl ether and finally stirred at 60 ° C. for a total of 54 hours. After the 8th and 42nd hour, 0.05 g of NaH and 0.4 g of diethylene glycol chloroethyl methyl ether are added again. After the mixture has cooled, it is poured onto water and extracted with ethyl acetate. After drying over MgSO₄ and evaporation of the extract, the compound (A17) is obtained, which is still chromatographed on silica gel with hexane / ethyl acetate (1: 1).

1 H-NMR (250 MHz, CDCl₃): 1.60 (s, C H ₃); 3.33 (s, OC H ₃); 5.47 (AB, C H₂ ); 5.93 [d, H -C (1 ')]; 7.65 [s, H -C (6)].
MS: 840 (M⁻).

Example A18

3.2 g of the product prepared in Example A17 is hydrogenated in 35 ml of tetrahydrofuran over 0.6 g of Pd / C (5%) at normal pressure and 22 ° C. After 1 hour (H₂ uptake: 96%), the hydrogenation mixture is filtered clear and evaporated. The residue is dissolved in 30 ml of methanol and mixed with 3.5 ml of a 30% solution of NaOCH₃ in methanol. After stirring at 20 ° C. for 24 hours, the reaction solution is poured onto water and extracted with ethyl acetate. After drying over MgSO₄ and evaporation of the extract, compound (A18) is obtained.

1 H-NMR (250 MHz, CDCl₃): 1.60 (s, C H ₃); 3.55 (s, OC H ₃); 5.96 [d, J = 2 Hz, H -C (1 ')]; 7.62 [s, H -C (6)]; 8.96 (s, NH ).
MS: 720/722/724 (M⁺).

Example A19

2.3 g of the product prepared in Example A18 are dissolved in 75 ml of methanol and hydrogenated at 35 ° C. with the addition of 1.15 g of sodium acetate and 1.0 g of Pd / C (5%). After 43 hours, the catalyst is filtered off and the filtrate is concentrated. The residue is filtered through silica gel to remove salts with ethyl acetate / methanol (4: 1). Compound (A19) is obtained.

1 H-NMR [250 MHz, methanol (D₄)]: 1.72 (s, C H ₃); 3.22 (s, OC H ₃); 5.80 [d, J = 5Hz, HC (1 ')]; 7.78 [s, H -C (6)].
MS: 404 (M).

Example A20

400 mg of the product prepared in Example A19 are dissolved in 8 ml of absolute pyridine. At 20 ° C., 0.34 g of DMTCl is added and the mixture is stirred for 20 hours. Then it is diluted with 30 ml of methylene chloride and poured onto water. The organic phase is washed a second time with water and then dried (MgSO₄) and evaporated. The residue is chromatographed on silica gel with ethyl acetate / triethylamine (97: 3). Compound (A20) is obtained.

1 H-NMR (250 MHz, CDCl₃; all signals are broad): 1.40 (s, C H ₃); 3.35 (s, OC H ₃); 3.70 (s, 2 x OC H ₃); 6.02 (s, HC (1 '); 7.66 [s, H -C (6)].
MS: 705 (MH).

Example A21

To introduce 0.12 g of diisopropylammonium tetrazolide and 0.20 g of cyanoethyltetraisopropylphosphoramidite in 2.5 ml of absolute methylene chloride, 0.39 g of the product prepared in Example A20 in 2.5 ml of methylene chloride is added dropwise. After stirring for 28 hours at 20 ° C, the reaction solution is poured onto a saturated aqueous NaHCO₃ solution and extracted with methylene chloride. The evaporated The residue is chromatographed on silica gel with ethyl acetate / triethylamine (97: 3). Compound (A21) is obtained.

1 H-NMR (250 MHz, CDCl₃): The protons of HC (1 ') appear as doublets at 6.02 and 6.07 ppm.
31 P NMR (CDCl3): 149.95 and 149.88 ppm

Example A22

21.4 g uracil are suspended in 250 ml dichloroethane. After adding 116 g of BSA, the mixture is heated to 80 °. Solution occurs after 30 minutes. It is cooled to 40 ° C. and 50.0 g of the product produced in Example A3, dissolved in 350 ml of dichloroethane, and 27.4 g of SnCl₄ are added. The solution is then kept at 80 ° C. for 5 hours. After cooling, it is poured onto saturated NaHCO₃ solution, the resulting 2-phase suspension is filtered and the organic phase is separated off. The aqueous phase is extracted again with CH₂Cl₂. The collected extracts are dried over MgSO₄ and evaporated. The residue is digested in acetonitrile. Compound (A22) is obtained.

NMR (250 MHz, CDCl₃): 2.22 (s, OAc), 5.35 (t, HC (2 ')), 6.07 (d, J = 4 Hz, HC (1')), 5.53 and 7.23 (each , J = 8 Hz, HC (5) and HC (6)), 8.57 (s, NH).
MS: 602

Example A23

To prepare 60 ml of THF and 1.47 g of NaH (100%), 35.6 g of the product prepared in Example A4, dissolved in 300 ml of THF, are added at 60 ° C. and 1 Hour further heated. Then 14.0 g of R - (-) glycidyl tosylate are added in portions. After 2.5 hours, 0.1 g of NaH is added again. After 4 hours of heating, the mixture is cooled, poured onto ice and extracted with ethyl acetate. The evaporated extract is chromatographed (silica gel, toluene / ethyl acetate 9: 1). Compound (A23) is obtained.

NMR (250 MHz, CDCl₃): 1.59 (s, CH₃), 5.48 (AB, CH₂), 5.92 (s, 1.5 Hz, HC (1 ')), 7.62 (s, HC (6)).
FAB-MS: 751 (M + H)

Example A24

5.1 g of the product prepared in Example A23 and 4.93 g of 1-hexadecanol are dissolved in 50 ml of CH₂Cl₂. 0.96 g of boron trifluoride diethyl etherate (d = 1.13 g / ml) are added. This solution is stirred for 19 hours at room temperature, then poured onto H₂O and extracted with CH₂Cl₂. The evaporated extract is chromatographed on silica gel (ethyl acetate / hexane 1: 1). Compound (A24) is obtained.

NMR (250 MHz, CDCl₃): 1.60 (s, CH₃), 5.47 (AB, CH₂), 5.93 (d, J = 1.5 Hz, HC (1 ')), 7.63 (s, HC (6) ).
FAB-MS: 1027 (M + Cl) ⁻

Example A25

3.45 g of the product prepared in Example A24 are heated in 60 ml of THF with 0.15 g of NaH (100%) for 45 minutes. Then 0.63 g of MeI are added and heating is continued for 1.5 hours. After cooling, the reaction mixture is poured onto water and extracted with ethyl acetate. The evaporated The residue is filtered through a short silica gel column in toluene. Compound (A25) is obtained.

NMR (250 MHz, CDCl₃): 1.59 (s, CH₃), 5.45 (AB, CH₂), 5.93 (d, J = 1.5 Hz, HC (1 ')), 7.63 (s, HC (6)).
MS (DCl): 1024 (M + NH₄) ⁺

Example A26

3.3 g of the product prepared in Example A25 are hydrogenated in 70 ml of THF over 0.7 g of Pd / C (33 hours, H₂ absorption 104%). The reaction mixture is filtered and evaporated. The residue is dissolved in 10 ml of THF and 10 ml of methanol and mixed with NaOCH₃ / HOCH₃ solution (30%). After stirring for 24 hours at room temperature, approximately half of the volume is concentrated, then poured onto water and extracted with ethyl acetate. The evaporated extract is chromatographed on silica gel (ethyl acetate / toluene 1: 2). Compound (A26) is obtained.

NMR (250 MHz, CDCl₃): 1.62 (s, CH₃), 3.40 (s, OCH₃), 5.98 (d, J = 2 Hz, HC (1 ')), 7.62 (s, HC (6)), 8.57 ( s, NH).
MS (DCl): 886

Example A27

1.21 g of the product prepared in Example A26 are hydrogenated with 50 ml of methanol and 5 ml of toluene over 0.2 g of Pd / C (10%) and 0.258 g of MgO (2 hours, H₂ uptake 220 ml). The reaction mixture is filtered, evaporated and filtered through silica gel (removal of Mg salts). Compound (A27) is obtained.

NMR (250 MHz, DMSO-D₆): 1.72 (s, CH₃), 5.05 (d, OH), 5.18 (t, OH), 5.84 (d, J = 6 Hz, HC (1 ')), 7.77 (s , HC (6)).
FAB-MS: 569 (MH).

Example A28

0.83 g of the product prepared in Example A28 are dissolved in 50 ml of pyridine and evaporated. The residue is mixed with 8 ml of pyridine and 0.6 g of DMT chloride and stirred for 44 hours at room temperature. The solution is poured onto H₂O and extracted with ethyl acetate. The evaporated residue is chromatographed on silica gel (toluene / ethyl acetate 1: 1 + 1% NEt₃ addition). Compound (A28) is obtained.

NMR (250 MHz, CDCl₃): 1.35 (s, CH₃), 3.47 (s, OCH₃), 3.80 (s, OCH₃), 6.03 (d, J = 4 Hz, HC (1 ')), 7.64 (s, HC (6)), 8.20 (s, NH).
FAB-MS: 873 (M + H).

Example A29

0.95 g of the product prepared in Example A28 are mixed with 10 ml of CH₂Cl₂, 0.245 g of diisopropylammonium tetrazolide and 0.39 g of cyanethyltetraisopropylphosphorodiamidite. After 3 days of stirring at room temperature, the reaction mixture is poured onto saturated NaHCO₃ solution and extracted with CH₂Cl₂. The evaporated residue is chromatographed on silica gel (toluene / ethyl acetate 2: 1 + 1% NEt₃ addition). Compound (A29) is obtained.

1 H-NMR (250 MHz, CDCl₃): 5.97 and 6.05 (each d, J = 4 Hz, each HC (1)), 7.62 and 7.70 (each s, each HC (6)).
P-NMR (250 MHz, CDCl₃): 150.012 and 150.102.
FAB-MS: 1071 (MH).

Example A30

15.6 g hypoxanthine and 46.6 g BSA are refluxed in 300 ml dichloroethane. Solution occurs after 30 minutes. 30.0 g of the product prepared in Example A3 and 16.5 g of TMS triflate are added and the mixture is boiled for a further 16 hours. After cooling, the reaction mixture is poured onto saturated NaHCO₃ solution and the resulting suspension is filtered. The filtrate is extracted with CH₂Cl₂. The organic phase is dried over MgSO₄ and evaporated. The residue is chromatographed (methanol / ethyl acetate 1: 1). Compound (A30) is obtained.

1 H-NMR (250 MHz, CDCl₃): 6.22 (d, 3 Hz, HC (1 ')), 8.14 and 8.37 (each s, HC (2) and HC (8)).
FAB-MS: 625 MH

Example A31

3.8 g of the product prepared in Example A30 are placed in 150 ml of CH₂Cl₂. A mixture of 4.8 ml SOCl₂ and 2.4 ml DMF is added dropwise while cooking. After 3 hours of boiling again 1.2 ml SOCl₂ and 0.6 ml DMF are added. After a total of 5.5 hours of boiling, the reaction mixture is reduced to a saturated NaHCO₃ solution entered. After stirring for 20 minutes, the mixture is extracted with CH₂Cl₂. The extract is dried over MgSO₄ and evaporated. Compound (A31) is obtained.

NMR (250 MHz, CDCl₃): 5.83 (dd, HC (2 ')), 6.33 (d, J = 3 Hz, HC (1')), 8.47 and 8.73 (each s, HC (2) and HC (8 )).
MS: 664

Example A32

30.4 g of the product prepared in Example A31 are dissolved in 200 ml of methanol and 100 ml of THF. 16.9 g of 30% NaOMe / methanol are added dropwise. After stirring for 3 hours, the mixture is poured onto H 2 O and extracted with ethyl acetate. The extract is dried over MgSO₄ and evaporated. Compound (A32) is obtained.

NMR (250 MHz, CDCl₃): 4.20 (s, OCH₃), 6.10 (d, J = 6 Hz, HC (1 ')), 8.17 and 8.48 (each s, HC (2) and HC (8)).
MS: 598

Example A33

1.17 g of NaH (100%) are added to 25.4 g of the product prepared in Example A32 in 250 ml of THF. 6.76 g of 2-bromoethyl methyl ether are added dropwise while boiling. After 6 hours of boiling again 0.5 g of NaH and 1.9 ml Bromide added. After 23 hours the reaction mixture is cooled and poured onto H₂O. After extraction with ethyl acetate, the mixture is evaporated and the residue is chromatographed. (Silica gel, hexane / acetone 2: 1). Compound (A33) is obtained.

1 H-NMR (250 MHz, CDCl₃): 3.27 (OCH₃), 4.20 (OCH₃), 6.27 (d, J = 5 Hz, HC (1 ')), 8.24 and 8.52 (each s, HC (2) and HC ( 8th)).
MS (DCI): 656

Example A34

2.2 g of the product prepared in Example A33 is hydrogenated in 45 ml of methanol with 0.66 g of Pd / C (10%) and with the addition of 402 ml of MgO. After 13 hours (H₂ intake: 485.5 ml H₂), the mixture is filtered clear and evaporated. Chromatography is carried out on silica gel to remove magnesium salts (ethyl acetate / methanol 9: 1). Compound (A34) is obtained.

NMR (250 MHz, DMSO (-d₆)): 4.4 (t, OH), 5.23 (OH), 5.94 (d, J = 6.5 Hz, HC (1 ')), 8.40 and 8.55 (each s, HC (2) and HC (8)).
FAB MS: 339 (MH) ⁻.

Example A35

2.46 g of the product prepared in Example A34 are dissolved in 80 ml of methanol and placed in an autoclave. 20 g of NH 3 are pressed on and the apparatus is kept at 120 ° C. for 12 hours (internal pressure: 17 bar). The reaction mixture is evaporated and chromatographed on silica gel (ethyl acetate / methanol). Compound (A35) is obtained.

NMR (250 MHz, DMSO-D₆): 5.32 (d, OH), 5.60 (t, OH), 6.15 (d, J = 6 Hz, HC (1 ')), 8.33 and 8.57 (each s, HC (2 ) and HC (8)).
MS (DCI): 326 (M + H).

Example A36

0.48 g of the product obtained in Example A35 are dissolved in 5 ml of methanol and 0.28 g of N-methyl-2,2-dimethoxypyrrolidine is added. After stirring for 18 hours, the mixture is evaporated. The residue is mixed twice with pyridine and evaporated. The remaining residue is dissolved in 10 ml of pyridine and stirred with 0.5 g of DMT chloride for 40 hours at room temperature. Then it is poured onto H₂O and extracted with CH₂Cl₂. The evaporated extract is chromatographed on silica gel (ethyl acetate / methanol 9: 1). Compound (A36) is obtained.

NMR (250 MHz, CDCl₃): 3.09 (s, NMe), 3.72 (s, OCH₃), 6.08 (d, J = 6 Hz, HC (1 ')), 7.98 and 8.43 (each s, HC (2) and HC (8)).
FAB MS: 709 (M + H)

Example A37

0.63 g of the trityl derivative obtained in Example A36 in 6 ml of CH₂Cl₂ are added to 0.38 g of cyanethyltetraisopropylphosphorodiamidite and 0.24 g of diisopropylammoniumtetrazolide in 6 ml of CH₂Cl₂. After stirring for 40 hours, the reaction mixture is poured onto saturated NaHCl₃ solution and extracted with CH₂Cl₂. The evaporated extract is chromatographed (ethyl acetate + 1% NEt₃). Compound (A37) is obtained.

31 P NMR (250 MHz, CDCl3): 149.8 and 150.4.
FAB-MS: 909 (M + H)

Example A38

7.1 g of the epoxide prepared in Example A23 is dissolved in 70 ml of THF, mixed with 0.43 g of NaBH₄ and stirred at room temperature. For this, 0.5 g of BF₃ · OEt₂ are added. After a total of 26 hours, the mixture is poured onto water and extracted with ethyl acetate. After evaporation, compound (A38) is obtained.

NMR (250 MHz, CDCl₃): 1.16 (d, J = 7.5 Hz, CH₃), 1.61 (s, CH₃), 5.48 (AB, CH₂), 5.92 (d, J = 1 Hz, HC (1 ') ), 7.69 (s, HC (6)).
FAB-MS: 787 (M + Cl) ⁻.

Example A39

6.9 g of the alcohol obtained in Example A38 are methylated with 1.56 g NaH (100%) and 1.56 g MeJ in 70 ml THF (60 ° C., 5 hours). After the usual work-up, the compound (A39) is obtained.

NMR (250 MHz, CDCl₃): 1.13 (d, J = 7.5 Hz, CH₃), 1.60 (s, CH₃), 3.33 (s, OCH₃), 5.49 (AB, CH₂), 5.95 (A, J = 2 Hz, HC (1 ')), 7.64 (s, HC (6)).
MS (DCI): 766 (M⁻).

Example A40

14.4 g of the derivative prepared according to Example A4 are heated to 60 ° C. in 150 ml THF and 0.65 g NaH (100%) for 30 minutes. The mixture is then cooled to 25 ° C. and 5.92 g of D-α, β-isopropylidene glycerol-γ-tosylate are added. After 1.5 hours, stirring is continued for 3 hours at 60 ° C. The cooled reaction mixture is poured onto water, extracted with ethyl acetate and evaporated. The residue is chromatographed on silica gel (toluene / ethyl acetate 4: 1). Compound (A40) is obtained.

NMR (250 MHz, CDCl₃): 1.34 and 1.41 (each s, each CH₃), 1.58 (s, CH₃), 5.48 (AB, CH₂), 5.92 (d, J = 1.5 Hz, HC (1 ')) , 7.68 (s, HC (6)).
MS (DCI): 843 (M + Cl) ⁻.

Example A41

3.13 g of the phosphoramidite prepared according to Example A13 are dissolved in 50 ml of acetonitrile. 9.4 ml of NEt₃ and 6.17 g of 1,2,4-triazole are added. 1.53 g of POCl₃ are then added dropwise with stirring in such a way that 30 ° C. are not exceeded. After stirring for 4 hours at room temperature, the reaction mixture is poured onto 1 N NaHCO 3 (150 ml) and extracted with ethyl acetate. The evaporated extract is dissolved in a little CH₂Cl₂ and stirred in 100 ml of n-pentane. The connection (A41) fails.

1 H-NMR (250 MHz, CDCl₃): 6.02 and 6.06 (each d, J = 1.5 Hz, HC (1 ')), 8.44 and 8.47 (each s, HC (6)), 8.07 and 9.35 (each s , triazole H).
31 P NMR (250 MHz, CDCl3): 149.433 and 150.253
MS (DCI): 869 (M⁻)

Example A42

8.88 g of N²-isobutyryl-O⁶-benzylguanine [prepared according to the analogous procedure by Jenny, TF, Schneider, KC, Benner, SA, Nucleosides and Nucleotides 11: 1257-1261 (1992)] and 34.8 g of N, O- Bis- (trimethylsilyl) acetamide are suspended in 130 ml of toluene and heated to 100 ° C. until solution occurs. 13.0 g of the ribose derivative prepared according to Example A3 and 6.33 g of trimethylsilyl trifluoromethanesulfonate are added at 50 ° C. The colorless solution is stirred for 4 hours at 100 ° C. and after cooling to 200 ml of saturated NaHCO₃ solution poured. After extraction with ethyl acetate, it is dried over MgSO₄ and evaporated. The residue is chromatographed on 1.2 kg of silica gel (toluene / ethyl acetate 4: 1). Compound (A42) is obtained.

1 H-NMR (250 MHz, CDCl₃): 1.26 (d, J = 7.5 Hz, CH₃), 2.23 (s, OAc), 5.61 (AB, OCH₂), 5.78 (dd, HC (2 ')), 6.19 (d, J = 3 Hz, HC (1 ')), 7.83 (s, NH), 8.00 (s, HC (8)).
MS (DCI): 801 (M⁻).

Example A43

7.4 g of the derivative prepared in Example A42, dissolved in 80 ml of methanol, are mixed with 0.83 g of a 30% NaOMe / methanol solution and stirred for 30 minutes at room temperature. The solution is poured onto H₂O and extracted with ethyl acetate. The extract is evaporated after drying over MgSO₄. Compound (A43) is obtained.

NMR (250 MHz, CDCl₃): 1.29 (d, J = 7.5 Hz, CH₃), 5.61 (AB, OCH₂), 5.99 (d, J = 6 Hz, HC (1 ')), 7.97 (s, NH ), 8.04 (s, HC (8)).
MS (DCI): 760 (M + H) ⁺

Example A44

630 mg NaH (100%) are added to 9.0 g of the compound prepared according to Example A43 in 100 ml THF and the mixture is stirred for 15 minutes. Then 1.97 g of 2-bromoethyl methyl ether are injected. After 25 hours, 2.0 g of bromide are added again. After stirring for a total of 48 hours, the mixture is poured into water and extracted with ethyl acetate. The evaporated extract is chromatographed on silica gel (toluene / ethyl acetate 4: 1). Compound (A44) is obtained.

NMR (250 MHz, CDCl₃): 3.28 (s, OCH₃), 5.56 (AB, OCH₂), 6.10 (d, J = 5 Hz, HC (1 ')), 7.90 (s, HC (8)).
FAB-MS: 748 (M + H) ⁺.

Example A45

Starting from compound (A3), further bases are introduced. The products are listed in Table 2.

Example B: Preparation of oligonucleotides

Using the dimethoxytritylated and 3'-activated [3 '- (β-cyanoethoxy-di (i-propylamino) phosphoramidite)] nucleosides or such naturally activated nucleosides, oligonucleotides are bound to a solid support (controlled pore glass, CPG) and the synthesis was carried out on a DNA synthesizer (Applied Biosystems, Model 380 B, standard phosphoramidite chemistry and iodine oxidation) according to the manufacturer's standard protocols [see also "Oligonucleotide synthesis a practical approach" MJ Gait; IRL Press 1984 (Oxford-Washington DC)]. After Coupling of the last nucleoside building block, the 5'-protected oligonucleotide is detached from the support by simultaneous cleavage of all other protective groups by treatment with concentrated aqueous ammonia and then purified using "reverse phase" HPLC using 50 mM ammonium acetate buffer (pH 7) / acetonitrile . The 5'-dimethoxytrityl protective group is then removed by treatment with 80% aqueous acetic acid for 20 minutes, the oligonucleotide is precipitated with ethanol and isolated by centrifugation. The purity of the oligonucleotide is checked by gel electrophoresis (polyacrylamide) and its identity by means of matrix-assisted laser desorption time-of-flight mass spectroscopy (MALDI-TOF MS).

Example C1: Affinity; Interaction of the oligonucleotides (antisense) with complementary oligoribonucleotide sequences (sense)

The interaction of the oligonucleotides with the corresponding base-complementary oligomers of the natural ribonucleotides is characterized by the recording of UV melting curves and the T _m values determined therefrom. This standard method is described, for example, by Marky, LA, Breslauer, KJ, Biopolymers 26: 1601-1620 (1987).
A solution of the oligonucleotides and the corresponding base-complementary natural oligoribonucleotides is prepared in 10 mM phosphate buffer, 100 mM NaCl, 0.1 mM EDTA, pH = 7.0 (c = 4 × 10 / M / oligonucleotide) and the change in absorbance recorded at 260 nm depending on the temperature (15 to 95 ° C). The T _m value is determined from the melting curves obtained (Table 3).

Example D2: Specificity; Interaction of the oligonucleotide with base-complementary oligoribonucleotides, in which a wrong nucleoside (Y) was incorporated

Solutions of the oligonucleotide are prepared with the corresponding base-complementary oligonucleotides of the sequences r (GGA CCG GAA YGG TAC GAG) in 10 mM phosphate buffer, 100 mM NaCl, 0.1 mM EDTA, pH 7, (c = 4 × 10⁻⁶ M / oligonucleotide ) and measures the change in absorbance at 260 nm depending on the temperature (15 ° C to 95 ° C). The T _m value is determined from the curves. The results are shown in Table 4.

Example C3: Nuclease stability; Enzymatic hydrolysis of various oligonucleotides of sequence d (TCC AGG TGT CCG ttt C)

14 µg each of the synthetic oligonucleotide or the corresponding natural oligomer are incubated in 200 µl 10% heat-inactivated serum from calf fetuses at 37 ° C (c = 70 µg / ml). After 0.5; 1; 2; 4; 6, 24 and 48 hours, 15 ul of the reaction solution are quenched by adding to 25 ul 9 M urea and trisborate buffer (pH 7) and stored at -20 ° C until measurement. The quenched reaction solutions are separated by means of polyacrylamide gel electophoresis and the cleavage products are determined via the phosphorus content (phospho-imagers method). The ratio R of the sum of the concentrations of the completely intact oligonucleotide (c _n ^(t) ) and the fragment (c _n-1 ^(t) ) resulting from cleavage of the natural C building block from the 3 'end at a given time t to the initial concentration of the completely intact oligonucleotide at time t = 0 (c _n ⁽⁰⁾ ) R = (c _n ^(t) + c _n-1 ^(t) ) / c _n ⁽⁰⁾ is plotted against time. The determined half-lives τ _1/2 - those are the times for which R = 0.5 -

Claims (62)

Compounds of formula I.

wherein R₁ and R₂ independently represent hydrogen or a protecting group or R₁ has these meanings and R₂ stands for a residue forming a phosphorus-containing nucleotide bridge group; B represents a purine or pyrimidine residue or an analog thereof; and R₃ represents a radical of formula Ia, Ib or Ic

wherein R₄ is hydrogen, C₁-C₂₁ alkyl, C₂-C₂₁ alkenyl, C₂-C₂₁ alkynyl or -C (= O) alkyl; R₅ is hydrogen, C₁-C₁₀ alkyl, -CH₂-O-R₆ or a radical of the formula Ib; R₆ is hydrogen, C₁-C₂₂-alkyl, C₃-C₂₁-alkenyl, partially or completely fluorine-substituted C₁-C₁₀-alkyl or - [(CH₂) ₂-O] _m -R₇; R₇ is hydrogen or C₁-C₂₁ alkyl; Z - (CH₂) _p - or - (CH₂-CH-O) _q -CH₂CH₂-, where Z in the case of -CH₂- is unsubstituted or with one or more C₁-C₁₀-alkyl, C₅-C₆-cycloalkyl or unsubstituted or with C₁-C₄ alkyl substituted phenyl may be independently substituted; n is a number from 1 to 12; m is a number from 1 to 4; p is a number from 1 to 10; and q represents a number from 1 to 4; where R₄ for the case n = 1 and R₅ = hydrogen is not hydrogen. Compounds according to claim 1, characterized in that R₁ and R₂ each represent hydrogen. Compounds according to claim 1, characterized in that R₁ and R₂ denote the same protective groups. Compounds according to Claim 1, in which B, as a purine radical or an analog thereof, represents a radical of the formula II, IIa, IIb, IIc, IId, IIe or IIf,

wherein R _{b1 represents} H, Cl, Br, OH or -O-C₁-C₁₂-alkyl, and R _b2 , R _b3 and R _b5 independently of one another H, OH, SH, NH₂, NHNH₂, NHOH, NHO-C₁-C₁₂ -Alkyl, -N = CH-N (C₁-C₁₂-alkyl) ₂, -N = CH-N-cycloalkyl, F, Cl, Br, C₁-C₁₂-alkyl, hydroxy-C₁-C₁₂-alkyl, Amino-C₁-C₁₂-alkyl, C₁-C₁₂-alkoxy, benzyloxy, C₁-C₁₂-alkylthio, where the hydroxyl and amino groups are unsubstituted or substituted with a protective group, phenyl, benzyl, primary amino with 1 to 20 C atoms or secondary amino with 2 to 30 carbon atoms, R _{b4 are} hydrogen, CN or -C≡CR _b7 and R _b6 and R _{b7 are} hydrogen or C₁-C₄-alkyl. Compounds according to claim 4, wherein the protective group for hydroxyl and amino groups is C₁-C₈ acyl. Compounds according to claim 4, wherein the primary amino contains 1 to 12 carbon atoms and the secondary amino contains 2 to 12 carbon atoms. Compounds according to claim 4, wherein the primary amino and secondary amino are radicals of the formula R _a1 R _a2 N, in which R _{a1 is} H or independently has the meaning of R _a2 , and R _{a2 is} C₁-C₂₀-alkyl, aminoalkyl , -Hydroxyalkyl; Carboxyalkyl or carbalkoxyalkyl, the carbalkoxy group containing 2 to 8 carbon atoms and the alkyl group 1 to 6, preferably 1 to 4 carbon atoms; C₂-C₂₀ alkenyl; Phenyl, mono- or di- (C₁-C₄ alkyl or alkoxy) phenyl, benzyl, mono- or di- (C₁-C₄ alkyl or alkoxy) benzyl; or 1,2-, 1,3- or 1,4-imidazolyl-C₁-C₆-alkyl, or R _a1 and R _a2 together tetra- or pentamethylene, 3-oxa-1,5-pentylene, -CH₂-NR _a3 -CH₂CH₂- or -CH₂CH₂-NR _a3 -CH₂CH₂-, in which R _{a3 represents} H or C₁-C₄-alkyl, the amino group in the aminoalkyl being unsubstituted or substituted by one or two C₁-C₄-alkyl or hydroxyalkyl groups, and the hydroxyl group in the hydroxyalkyl is optionally etherified with C₁-C₄-alkyl. Compounds according to claim 6, wherein the primary amino and secondary amino are methyl, ethyl, dimethyl, diethyl, allyl, mono- or di (hydroxyeth-2-yl), phenyl and benzyl, acetyl -, Isobutyryl and Benzoylamino acts. Compounds according to claim 4, wherein R _b1 in formulas II, IIb, IIc, IId and IIe represents hydrogen. Compounds according to claim 4, wherein R _b5 in formula IId represents hydrogen. Compounds according to claim 4, wherein R _b2 and R _b3 in the formulas II, IIa, IIb, IIc, IId and IIf independently of one another H, F, Cl, Br, OH, SH, NH₂, NHOH, NHNH₂, methylamino, dimethylamino, benzoylamino , Methoxy, ethoxy and methylthio. Compounds according to Claim 4, in which B is a purine residue or a residue of a purine analog from the series adenine, N-methyladenine, N-benzoyladenine, 2-methylthioadenine, 2-aminoadenine, 6-hydroxypurine, 2-amino-6-chloropurine, 2-amino -6-methylthiopurine, guanine and N-isobutyrylguanine. Compounds according to Claim 1, in which B in formula I as pyrimidine residue represents an uracil, thymine or cytosine residue of the formula III, IIIa, IIIb or IIIc,

wherein R _b6 H or C₁-C₄-alkyl and R _b8 H, OH, SH, NH₂, NHNH₂, NHOH, NHO-C₁-C₁₂-alkyl, -N = CH-N (C₁-C₁₂-alkyl) ₂, -N = CH-N-cycloalkyl, F, Cl, Br, C₁-C₁₂-alkyl, hydroxy-C₁-C₁₂-alkyl, amino-C₁-C₁₂-alkyl, C₁-C₁₂-alkoxy, benzyloxy, C₁-C₁₂-alkylthio, where the hydroxyl and amino groups are unsubstituted or substituted by a protective group, phenyl, benzyl, primary amino having 1 to 20 carbon atoms, secondary amino having 2 to 30 carbon atoms, C₁-C₁₂ alkenyl or C₁-C₁₂ alkynyl, and the NH₂ group in formula IIIb is unsubstituted or substituted by C₁-C₆-alkyl, benzoyl or a protective group, and the dihydro derivatives of the radicals of the formulas III, IIIa, IIIb and IIIc. Compounds according to claim 13, wherein R _{b8 is} H, C₁-C₆ alkyl or hydroxyalkyl, C₂-C₆ alkenyl or alkynyl, F, Cl, Br, NH₂, benzoylamino, mono- or di-C₁-C₆ alkylamino. Compounds according to claim 13, wherein R _{b8 is} H, C₁-C₆ alkyl or alkoxy or hydroxyalkyl, C₂-C₆ alkenyl or alkynyl, F, Cl, Br, NH₂, benzoylamino, mono- or di-C₁-C₆-alkylamino . Compounds according to Claim 14, in which R _{b8 is} H, F, Cl, Br, NH₂, NHCH₃, N (CH₃) ₂, C₁-C₄-alkyl, C₂-C₄-alkenyl- (1) or C₂-C₄-alkynyl- (1 ) means. Compounds according to claim 15, wherein R _{b8 is} H, C₁-C₄-alkyl, C₂-C₄-alkenyl- (1), C₂-C₄-alkynyl- (1), NH₂, NHCH₃ or (CH₃) ₂N. Compounds according to Claim 1, in which B as the residue of a pyrimidine analog is derived from uracil, thymine, cytosine, 5-fluorouracil, 5-chlorouracil, 5-bromouracil, dihydrouracil, 5-methylcytosine, 5-propinthymine and 5-propyne cytosine. Compounds according to Claim 1, characterized in that R₂ as a phosphorus-containing radical of the formula P1 or P2 forming a nucleotide bridge group

corresponds to, wherein Y _{a is} hydrogen, C₁-C₁₂-alkyl, C₆-C₁₂-aryl, C₇-C₂₀-aralkyl, C₇-C₂₀- alkaryl, -OR _b , -SR _b , -NH₂, primary amino, secondary ^amino , O ^⊖ M ^⊕ or S ^{⊖ represents} M ^⊕ ;
X _{a represents} oxygen or sulfur; R _{a is} hydrogen, M ^⊕ , C₁-C₁₂-alkyl, C₂-C₁₂-alkenyl, C₆-C₁₂-aryl, or the group R _a O- for N-heteroaryl-N-yl with 5 ring members and 1 to 3 N atoms stands; R _{b represents} hydrogen, C₁-C₁₂ alkyl or C₆-C₁₂ aryl; and M ^{⊕ represents} Na ^⊕ , K ^⊕ , Li ^⊕ , NH₄ ^⊕ or represents primary, secondary, tertiary or quaternary ammonium; where alkyl, aryl, aralkyl and alkaryl in Y _a , R _a and R _b is unsubstituted or substituted with alkoxy, alkylthio, halogen, -CN, -NO₂, phenyl, nitrophenyl, or halophenyl. Compounds according to claim 19, characterized in that R _{a is} β-cyanoethyl and Y _{a is} di (i-propyl) amino. Compound according to claim 1, characterized in that R₃ represents a radical of formula Ia. Compounds according to claim 21, characterized in that n stands for a numerical value from 1 to 8. Compounds according to claim 22, characterized in that n stands for a numerical value from 1 to 6. Compounds according to claim 21, characterized in that R₄ is hydrogen, C₁-C₂₁-alkyl, C₂-C₂₁-alkenyl or C₂-C₂₁-alkynyl. Compounds according to claim 24, characterized in that R₄ is hydrogen or C₁-C₄ alkyl. Compounds according to claim 21, characterized in that R₅ is hydrogen, C₁-C₅-alkyl or -CH₂-OH, -CH₂-O-C₁-C₂₂-alkyl or -CH₂-O - [(CH₂) ₂-O) _m -C₁ -C₁₀-alkyl, wherein m is a number from 1 to 4. Compounds according to claim 26, characterized in that R₅ is hydrogen, methyl or -CH₂-OH, -CH₂-O-C₁-C₂₂-alkyl or -CH₂-O - [(CH₂) ₂-O] _m -C₁-C₁₀-alkyl , wherein m is a number from 1 to 4. Compounds according to claim 1, characterized in that R₃ represents a radical of formula Ib. Compounds according to claim 28, characterized in that Z is -CH₂- or substituted -CH₂-. Compounds according to claim 1, characterized in that R₃ represents a radical of formula Ic. Process for the preparation of compounds of formula I,

wherein R₁ and R₂ independently represent hydrogen or a protecting group or R₁ has these meanings and R₂ stands for a residue forming a phosphorus-containing nucleotide bridge group; B represents a purine or pyrimidine residue or an analog thereof; and (a) R₃ is a radical of the formula Ia

wherein R₄ is hydrogen, C₁-C₂₁ alkyl, C₂-C₂₁ alkenyl, C₂-C₂₁ alkynyl or -C (= O) alkyl; R₅ is hydrogen, C₁-C₁₀ alkyl, -CH₂-O-R₆ or a radical of the formula Ib; R₆ is hydrogen, C₁-C₂₂-alkyl, C₃-C₂₁-alkenyl, partially or completely fluorine-substituted C₁-C₁₀-alkyl or - [(CH₂) ₂-O] _m , -R₇; R₇ is hydrogen or C₁-C₂₁ alkyl; n is a number from 1 to 12; and m represents a number from 1 to 4; where R₄ is not hydrogen for the case n = 1 and R₅ = hydrogen,
characterized in that a compound of formula IVa

wherein R₁₄ and R₁₅ represent the same or different protective groups and B is a purine or pyrimidine radical or an analog thereof, functional groups in the base radical B being protected by protective groups, reacted in an inert solvent with a compound of the formula A.

wherein R₄, R₅ and n have the meanings given above and X is Cl, Br, I, tosyl-O or mesyl-O; (b) R₃ is a radical of the formula Ic

characterized in that reacting a compound of formula IVa in an inert solvent with a compound of formula B.

wherein X is Cl, Br, I, Tosyl-O or Mesyl-O; (c) R₃ is a radical of the formula Ib

wherein R₅ is hydrogen, C₁-C₁₀ alkyl, -CH₂-O-R₆ or a radical of the formula Ib; R₆ is hydrogen, C₁-C₂₂-alkyl, C₃-C₂₁-alkenyl, partially or completely fluorine-substituted C₁-C₁₀-alkyl or - [(CH₂) ₂-O] _m -R₇; R₇ is hydrogen or C₁-C₂₁ alkyl; Z - (CH₂) _p - or - (CH₂-CH-O) _q -CH₂CH₂-, where Z in the case of -CH₂- is unsubstituted or with one or more C₁-C₁₀-alkyl, C₅-C₆-cycloalkyl or unsubstituted or with C₁-C₄ alkyl substituted phenyl may be independently substituted; m is a number from 1 to 4; p is a number from 1 to 10; and q represents a number from 1 to 4;
characterized in that the epoxide obtained in (b) is opened in the presence of H₂O and BF₃ and with a compound of the formula X '- (CH₂) qX' or X '- (CH₂-CH-O) _r -X', wherein X 'denotes Cl, Br, I, Tosyl-O or Mesyl-O, closes to a new ring; (d) R₃ represents a radical of the formula Ia,
characterized in that the epoxide obtained in (b) is reacted in the presence of R₆OH and BF₃ and optionally the released hydroxyl group is converted into an ether or ester; (e) R₃ represents a radical of the formula Ia,
characterized in that the epoxide obtained in (b) is hydrogenated in the presence of BF₃ and, for example, NaBH₄ and, if appropriate, the hydroxyl group released is converted into an ether or ester; (f) R₃ represents a radical of the formula Ia,
characterized in that the epoxide obtained in (b) is reacted with a corresponding Grignard reagent and, if appropriate, the hydroxyl group released is converted into an ether or ester; or (g) R₃ represents a radical of the formula Ia, Ib or Ic,
characterized in that a compound of formula IVb

wherein R₁₄ and R₁₅ have the meaning mentioned above and A represents a leaving group, substituted on the 2'-OH group by one of the processes described in (a) to (f) and introduces the base radical B by substitution; optionally splitting off the protective groups R₁₄ and R₁₅ and introducing the residue forming the phosphorus-containing nucleotide bridge group. Use of the compounds according to claim 1 for the production of oligonucleotides, the same or different monomer units of compounds of the formula I or the at least one monomer unit of compounds of the formula I in combination with monomer units of other natural or synthetic nucleosides, the oligonucleotides consisting of 2 to 200 monomer units. Use according to claim 32 for the production of oligonucleotides with 2 to 100 monomer units. Use according to claim 33 for the production of oligonucleotides with 2 to 50 monomer units. Use according to claim 34 for the production of oligonucleotides with 4 to 30 monomer units. Use according to claim 32 for the production of oligonucleotides with identical or different monomer units of compounds of the formula I. Use according to claim 32 for the production of oligonucleotides with the same monomer units of compounds of the formula I and monomer units of natural or synthetic nucleosides. Oligonucleotides of formula V

5'-U- (OYOV-) _x OYOW-3 '(V),

wherein x is a number from 0 to 200 and Y is a nucleotide bridge group; U, V and W each represent the same or different radicals of natural or synthetic nucleosides and at least one of the radicals U, V and / or W represents a radical of the formula VI

wherein B represents a purine or pyrimidine residue or an analog thereof; and R₃ represents a radical of formula Ia, Ib or Ic

wherein R₄ is hydrogen, C₁-C₂₁ alkyl, C₂-C₂₁ alkenyl, C₂-C₂₁ alkynyl or -C (= O) alkyl; R₅ is hydrogen, C₁-C₁₀ alkyl, -CH₂-O-R₆ or a radical of the formula Ib; R₆ is hydrogen, C₁-C₂₂-alkyl, C₃-C₂₁-alkenyl, partially or completely fluorine-substituted C₁-C₁₀-alkyl or - [(CH₂) ₂-O] _m -R₇; R₇ is hydrogen or C₁-C₂₁ alkyl; Z - (CH₂) _p - or - (CH₂-CH-O) _q -CH₂CH₂-, where Z in the case of -CH₂- is unsubstituted or with one or more C₁-C₁₀-alkyl, C₅-C₆-cycloalkyl or unsubstituted or with C₁-C₄ alkyl substituted phenyl may be independently substituted; n is a number from 1 to 12; m is a number from 1 to 4; p is a number from 1 to 10; and q represents a number from 1 to 4. Oligonucleotides according to claim 38, characterized in that the bridge group Y is -P (O) O ^⊖ -, -P (O) S ^⊖ -, -P (S) S ^⊖ -, -P (O) R₁₆- , -P (O) NR₁₇R₁₈-, or -CH₂-, in which R₁₆ represents H or C₁-C₆-alkyl, and R₁₇ and R₁₈ independently of one another have the meaning of R₁₆. Oligonucleotides according to claim 38, characterized in that the bridge group Y is -P (O) O ^⊖ -. Oligonucleotides according to claim 38, characterized in that x stands for a number from 0 to 100. Oligonucleotides according to claim 41, characterized in that x stands for a number from 1 to 50. Oligonucleotides according to claim 42, characterized in that x stands for a number from 3 to 29. Oligonucleotides according to claim 38, characterized in that the residues of formula VI are terminal and / or bound in the nucleotide sequence. Oligonucleotides according to claim 38, characterized in that the residues of formula VI are bound between residues of natural or synthetic nucleosides. Oligonucleotides according to claims 44 and 45, characterized in that 2 to 5 identical or different radicals of the formula VI follow one another. Oligonucleotides according to claim 38, characterized in that a total of 4 to 30 nucleoside units and 1 to 12 residues of the formula VI are contained. 38. Oligonucleotides according to claim 38, wherein B as a purine residue or an analog thereof represents a residue of the formulas II, IIa, IIb, IIc, IId, IIe or IIf,

wherein R _{b1 represents} H, Cl, Br, OH or -O-C₁-C₁₂-alkyl, and R _b2 , R _b3 and R _b5 independently of one another H, OH, SH, NH₂, NHNH₂, NHOH, NHO-C₁-C₁₂ -Alkyl, -N = CH-N (C₁-C₁₂-alkyl) ₂, -N = CH-N-cycloalkyl, F, Cl, Br, C₁-C₁₂-alkyl, hydroxy-C₁-C₁₂-alkyl, amino-C₁ -C₁₂-alkyl, C₁-C₁₂-alkoxy, benzyloxy, C₁-C₁₂-alkylthio, where the hydroxyl and amino groups are unsubstituted or substituted with a protective group, phenyl, benzyl, primary amino with 1 to 20 C atoms or secondary amino with 2 to 30 carbon atoms mean R _{b4 is} hydrogen, CN or -C≡CR _b7 , R _b6 and R _{b7 are} hydrogen or C₁-C₄ alkyl. 38. Oligonucleotides according to claim 38, wherein B represents an uracil, thymine or cytosine residue of the formula III, IIIa, IIIb or IIIc,

wherein R _b6 H or C₁-C₄-alkyl and R _b8 H, OH, SH, NH₂, NHNH₂, NHOH, NHO-C₁-C₁₂-alkyl, -N = CH-N (C₁-C₁₂-alkyl) ₂, -N = CH-N-cycloalkyl, F, Cl, Br, C₁-C₁₂-alkyl, hydroxy-C₁-C₁₂-alkyl, amino-C₁-C₁₂-alkyl, C₁-C₁₂-alkoxy, benzyloxy, C₁-C₁₂-alkylthio, the hydroxyl and amino groups being unsubstituted or substituted by a protective group, Phenyl, benzyl, primary amino with 1 to 20 C atoms, secondary amino with 2 to 30 C atoms, C₁-C₁₂ alkenyl or C₁-C₁₂ alkynyl, and the NH₂ group in formula IIIb unsubstituted or with C₁-C₆- Alkyl, benzoyl or a protective group is substituted, as well as the dihydro derivatives of the radicals of the formulas III, IIIa, IIIb and IIIc. Oligonucleotides according to claim 38, wherein R₃ represents a radical of formula Ia. Oligonucleotides according to claim 50, characterized in that n stands for a numerical value from 1 to 8. Oligonucleotides according to claim 51, characterized in that n stands for a numerical value from 1 to 6. Oligonucleotides according to claim 50, characterized in that R₄ is hydrogen, C₁-C₂₁-alkyl, C₂-C₂₁-alkenyl or C₂-C₂₁-alkynyl. Oligonucleotides according to claim 53, characterized in that R₄ is hydrogen or C₁-C₄ alkyl. Oligonucleotides according to claim 50, characterized in that R₅ is hydrogen, C₁-C₅-alkyl or -CH₂-OH, -CH₂-O-C₁-C₂₂-alkyl or -CH₂-O - [(CH₂) ₂-O] _m -C₁ -C₁₀-alkyl, wherein m is a number from 1 to 4. Oligonucleotides according to claim 55, characterized in that R₅ is hydrogen, methyl or -CH₂-OH, -CH₂-O-C₁-C₂₂-alkyl or -CH₂-O - [(CH₂) ₂-O] _m , -C₁-C₁₀- alkyl, in which m is a number from 1 to 4. Oligonucleotides according to claim 38, characterized in that R₃ represents a radical of the formula Ib. Oligonucleotides according to claim 57, characterized in that Z is -CH₂- or substituted -CH₂-. Oligonucleotides according to claim 38, characterized in that R₃ represents a radical of formula Ic. Use of the oligonucleotides of the formula V as diagnostics for the detection of viral infections or genetic diseases. Oligonucleotides of formula V for use in a therapeutic method for the treatment of diseases in warm-blooded animals, including humans, by interaction with nucleotide sequences in the body. Pharmaceutical preparation containing an effective amount of a nucleoside of the formula I or an oligonucleotide of the formula V alone or together with other active ingredients, a pharmaceutical carrier material and optionally auxiliary substances.